Microfluidic device for assessing material implanted fibrosis and cancer

Abstract

Fibrosis diagnostic systems, subsystems, and components thereof are provided. A fibrosis diagnostic system may include a microfluidic device. The fibrosis diagnostic system may be preloaded with one or more components. For example, the fibrosis diagnostic system may comprise one or more agents and a target object intended for implantation, such as tissue or a medical device, or both. The fibrosis diagnostic system may include one or more additional components and subsystems, such as a detector including a sensor configured to detect fibrosis of the target subject, a thermal subsystem, a fluid subsystem, a processor, or a user interface, or any combination thereof. Kits comprising one or more components of a fibrosis diagnostic system are provided. Screening methods for identifying therapeutic agents using a fibrosis diagnostic system or components thereof are provided. A non-transitory computer-readable medium storing a program is further provided.

Description

[0001] Cross-reference to related applications

[0002] This is an international application under the Patent Cooperation Treaty, which claims priority to U.S. Provisional Patent Application No. 63 / 335,527 filed on April 27, 2022, the contents of which are incorporated herein by reference in their entirety. Technical Field

[0003] The present disclosure relates to systems, devices, and methods for analyzing fibrosis associated with foreign matter and treatment methods based on the analysis. Background Art

[0004] Foreign body responses lead to fibrosis of implants and cancer tumors, which may cause multiple negative effects, such as hypertrophic scars, excessive inflammation, and reduced efficacy of cancer therapeutics. During the fibrosis formation process, crosstalk, primarily between macrophages and fibroblasts, leads to the separation of biomaterials from native tissues. Traditionally, these interactions have been evaluated by placing foreign bodies in vivo in mouse or rat models. Available in vitro models attempt to simulate scars including associated fibrosis. These models can provide relevant data, but cannot provide adequate models of the interactions that occur in humans, and therefore may have limited predictive value.

[0005] The present disclosure provides solutions to these technical problems. Summary of the invention

[0006] The present disclosure provides a fibrosis diagnosis system, a subsystem and its components. A fibrosis diagnosis system may include a microfluidic device. The microfluidic device may include a device housing, the device housing including a plurality of channels within the device interior, a plurality of diaphragms separating the channels from each other, and a plurality of channel ports in the device housing in fluid communication with the plurality of channels. A plurality of channels may include, for example, a first channel, a second channel and a third channel. The first channel may be a central channel, and the second channel and the third channel may be lateral channels on opposite sides of the central channel. The first channel may be configured to receive a target object, a matrix and a first biological composition, the first biological composition being configured to promote fibrosis of the target object. The second channel may be configured to receive a second biological composition, the second biological composition being configured to promote fibrosis of the target object. The first diaphragm may separate the first channel from the second channel. The third channel may be configured to receive a third biological composition, the third biological composition being configured to promote fibrosis of the target object. The second diaphragm may separate the first channel from the third channel. A plurality of channel ports may include any number of channel ports with a minimum of two channel ports. A plurality of channel ports may include one or more groups of channel ports. The fibrosis diagnosis system can be configured to form a gradient of at least one type of molecule across the first channel, the second channel or the third channel or any combination thereof. The fibrosis diagnosis system can be preloaded with one or more components. For example, the fibrosis diagnosis system can include a matrix, a first biological composition, a second biological composition or a third biological composition or any combination thereof. The fibrosis diagnosis system can include a target object. The fibrosis diagnosis system can include one or more additional components and subsystems, such as a detector, and the detector includes a sensor, a thermal subsystem, a fluid subsystem, a processor or a user interface configured to detect the fibrosis of the target object or any combination thereof. A kit comprising one or more components of the fibrosis diagnosis system is provided. A screening method using a fibrosis diagnosis system or its components is provided. The screening method includes, for example, a method for screening an anti-fibrotic agent or an anticancer agent. A non-transitory computer-readable medium is further provided, which stores a program containing instructions, and the instructions are used to perform the method of the present disclosure or to control the system of the present disclosure. BRIEF DESCRIPTION OF THE DRAWINGS

[0007] For a further understanding of the nature, objects and advantages of the present disclosure, reference should be made to the following detailed description, read in conjunction with the following drawings, wherein like reference numerals represent like elements.

[0008] Figure 1A is a perspective view of a microfluidic device according to the present disclosure.

[0009] Figure 1B yes Figure 1A A plan view of the microfluidic device shown in FIG.

[0010] Figure 2A is a perspective view of a fibrosis diagnostic system showing a microfluidic device detached from a detector according to the present disclosure.

[0011] Figure 2B FIG. 2 is a diagram showing a microfluidic device coupled to a detector according to the present disclosure. Figure 2A A perspective view of the fibrosis diagnostic system.

[0012] Figure 3 is a plan view of a microfluidic device according to the present disclosure.

[0013] Figure 4 is a plan view of a microfluidic device according to the present disclosure.

[0014] Figure 5 is a plan view of a microfluidic device according to the present disclosure.

[0015] Fig. 6A is a top perspective view of a microfluidic device according to the present disclosure.

[0016] Figure 6B yes Fig. 6A Front perspective view of the microfluidic device shown in .

[0017] Figure 6C yes Fig. 6A An enlarged perspective view of a portion of the microfluidic device shown in FIG.

[0018] Fig.6D yes Fig. 6A An enlarged plan view of a portion of the microfluidic device shown in FIG.

[0019] Fig. 6E yes Fig.6D A further enlarged plan view of a portion of the enlarged view shown in .

[0020] Fig. 7A is a schematic cross-sectional view of a loaded microfluidic device at a first point in time according to the present disclosure.

[0021] Figure 7B At the second time point Fig. 7A Schematic cross-sectional view of the loaded microfluidic device shown in .

[0022] Figure 8 is a schematic diagram of a fibrosis diagnostic system according to the present disclosure.

[0023] Fig. 9 is a schematic diagram of a fibrosis diagnostic system according to the present disclosure.

[0024] Fig.10 is a schematic diagram of a fibrosis diagnostic system according to the present disclosure.

[0025] Fig.11 , parts A, B and C show magnified images of cells stained with picrosirius red according to the present disclosure.

[0026] Fig.12 Shown are magnified fluorescent images of cells grown in 96-well plates uncoated or coated with fibronectin, with L-ascorbic acid, with or without TGF-β1, and fixed with 4% paraformaldehyde (PFA) in accordance with the present disclosure.

[0027] Fig.13 Shown are magnified fluorescent images of cells grown in 96-well plates uncoated or coated with fibronectin, with L-ascorbic acid, with or without TGF-β1, and fixed with methanol in accordance with the present disclosure.

[0028] Fig.14 Depicted are graphs of quantitative fluorescence intensity of collagen I and α-SMA channels at 4X and 10X magnification of cells grown in accordance with the present disclosure.

[0029] Fig.15 Depicted are graphs of quantitative fluorescence intensity of collagen I and α-SMA channels at 20X and 40X magnification of cells grown in accordance with the present disclosure.

[0030] Fig.16 Shown are magnified fluorescent images of cells grown in eight chamber glass slides uncoated or coated with fibronectin, with L-ascorbic acid, with or without TGF-β1, and fixed with 4% paraformaldehyde (PFA) in accordance with the present disclosure.

[0031] Fig.17 Shown are magnified fluorescent images of cells grown in eight chamber glass slides uncoated or coated with fibronectin, with L-ascorbic acid, with or without TGF-β1, and fixed with methanol in accordance with the present disclosure. DETAILED DESCRIPTION

[0032] The present disclosure provides a fibrosis diagnosis system. The fibrosis diagnosis system may include a microfluidic device. The microfluidic device may include a device housing, wherein the device housing includes a plurality of channels inside the device, inside the device, a plurality of diaphragms separating the channels from each other, and a plurality of channel ports in the device housing that are in fluid communication with the plurality of channels. A plurality of channels may include any number of channels with a minimum of two channels. A plurality of channels may be, for example, 2, 3, 4, 5, 6, 7, 8, 9 or 10 channels, or more than 10 channels. The number of diaphragms may be, for example, one less than the number of channels. For example, a plurality of channels may include a first channel, a second channel and a third channel having a first diaphragm and a second diaphragm. The number of diaphragms may be, for example, 1, 2, 3, 4, 5, 6, 7, 8 or 9 diaphragms, or more than 9 diaphragms. The first channel may be a central channel, and the second channel and the third channel may be lateral channels on opposite sides of the central channel (first channel). The first channel can be configured to receive a target object, a matrix and a first biological composition, the first biological composition being configured to promote fibrosis of the target object. The second channel can be configured to receive a second biological composition, the second biological composition being configured to promote fibrosis of the target object. The first diaphragm can separate the first channel from the second channel. The first diaphragm can include a first plurality of openings that provide fluid communication between the first channel and the second channel. The third channel can be configured to receive a third biological composition, the third biological composition being configured to promote fibrosis of the target object. The second diaphragm can separate the first channel from the third channel. The second diaphragm can include a second plurality of openings that provide fluid communication between the first channel and the third channel. A plurality of channel ports in the device housing that are in communication with a plurality of channel fluids can include any number of channel ports with a minimum of two channel ports. The plurality of channel ports can include one or more groups of channel ports, wherein a group of channel ports belongs to a particular channel. For example, a plurality of channel ports can include a first group of channel ports in communication with the first channel fluid, a second group of channel ports in communication with the second channel fluid, and a third group of channel ports in communication with the third channel fluid, wherein each group of channel ports includes one or more channel ports.

[0033] The fibrosis diagnostic system can be configured to form a gradient of at least one type of molecule across the first channel, the second channel or the third channel or any combination thereof. For example, at least one type of molecule can include a chemotactic molecule. The chemotactic molecule can promote the movement of at least one cell type present in the first biological composition, the second biological composition or the third biological composition or any combination thereof. At least one type of molecule can include a growth factor. The growth factor can promote the growth of a cell colony, and does so to form a gradient that includes the cell colony. The first biological composition, the second biological composition or the third biological composition or any combination thereof can include at least one type of molecule.

[0034] A plurality of channel ports may include any number of channel ports with a minimum of two channel ports. A channel may share one or more channel ports. A channel may have one or more dedicated channel ports. A plurality of channel ports may include at least one channel port for each channel. A plurality of channel ports may include at least two channel ports for at least one channel. A plurality of channel ports may include one or more groups of channel ports, wherein a group of channel ports belongs to a particular channel. For example, the second group of channel ports and the third group of channel ports each include two or more channel ports. Each group of channel ports may include any desired number of channel ports, for example, at least one channel port, two or more channel ports, three or more channel ports, or four or more channel ports. The second group of channel ports and the third group of channel ports each include a first channel port and a second channel port, for example, the first channel port is close to the first end of its corresponding channel, and the second channel is close to the second end of its corresponding channel. Each group of channel ports in the plurality of groups of channel ports may include a first channel port and a second channel port, for example, the first channel port is close to the first end of its corresponding channel, and the second channel is close to the second end of its corresponding channel. The first channel may include a central channel port, and the central channel port is configured to receive a target object into the first channel. The central channel port can be between the first end of the first channel and the second end of the second channel. For example, the central channel port can be equidistant between the first end and the second end (the end of the first channel). One or more channel ports in a plurality of channel ports can include seals, valves or closures or any combination thereof, and the seals, valves or closures or any combination thereof are configured to move between an open state and a closed state. One or more channel ports in a plurality of channel ports can be configured to be connected to a fluid subsystem, to transfer one or more components to a microfluidic device, or to transfer out or both from a microfluidic device. A channel port can be configured as an inlet or an outlet or both.

[0035] One or more retaining elements may be located in one or more channels of a multifluidic device. For example, one or more retaining elements may be located in one or more channels of a first channel, a second channel, and a third channel. The retaining element may be configured to retain at least one component that can be inserted into a channel of a microfluidic device, such as one or more channels of a first channel, a second channel, and a third channel. The retaining element may have any desired design or combination of designs. The retaining element may be permanent or removable. The retaining element may be movable within a microfluidic device or may be moved into or out of a microfluidic device. The retaining element may be retractable. The retaining element may include, for example, a modification of a channel surface. The modification may include, for example, a biomolecule. The biomolecule may include a polypeptide, a polysaccharide, a lipid, or a polynucleotide, or any combination thereof. The modification may include, for example, a polymer, a metal, or a ceramic, or any combination thereof. The device housing may include, for example, a polymer, a metal, or a ceramic, or any combination thereof. For example, the device housing may include a first polymer, and the modification may include a second polymer. The first polymer and the second polymer may differ from each other in chemical composition. The modification may include, for example, a physical structure. The physical structure and the device housing may include a common material. The modification may include a container for receiving a target object. Container can comprise the recess in the channel surface for example.Container, recess or other physical structure can have any desired geometry or size or both.For example, container, recess or physical structure can be circular, oval, triangular, square, rectangular, pentagonal, hexagonal, heptagonal, octagonal or trapezoidal or its any combination.The width, diameter, height or depth of container, recess or other physical structure or its any combination can be less than about 1.0 μm, about 1.0 μm to about 1.0cm, about 10 μm to about 100mm, about 50 μm to about 50mm, about 50 μm to about 5.0mm, about 50 μm to about 9.5mm, about 100 μm to about 1.0mm or greater than 1.0cm.Physical structure can comprise multiple physical structures.For example, multiple physical structures can comprise multiple recesses in the channel surface or multiple projections or both from the channel surface.

[0036] Microfluidic device and any component thereof can have any desired geometry, shape, size, dimension or relative proportion or any combination thereof.For example, length or width or two microfluidic devices can be less than 1.0mm, about 1.0mm to about 10cm, about 2.5mm to about 5.0cm, about 5.0mm to about 2.5cm, about 7.5mm to about 1.0cm or greater than 10cm.The thickness of microfluidic device can be less than about 0.001mm, about 0.001mm to about 1.0cm, about 0.01mm to about 0.5cm, about 0.1mm to about 0.1cm, about 1.0mm to about 0.5cm, about 10mm to about 100mm or greater than 1.0cm.The first channel, the second channel or the third channel or any combination thereof can include a curved cross section or a straight cross section or both.The curved cross section can be circular or elliptical.The first channel, the second channel or the third channel or any combination thereof can include a constant width or a constant diameter or both.The first channel, the second channel or the third channel or any combination thereof can include a variable width or a constant diameter or both. The width, diameter or height of passage or any combination thereof can be less than about 1.0 μm, about 1.0 μm to about 1.0 cm, about 10 μm to about 0.5 cm, about 50 μm to about 1.0 cm, about 100 μm to about 0.1 cm, about 1.0 mm to about 500 mm, about 10 mm to about 100 mm, or greater than 1.0 cm, or any value or any intermediate range therebetween. Two or more passages in a plurality of passages can have a common length. Passage length or width or both can be less than 1.0 mm, about 1.0 mm to about 10 cm, about 2.5 mm to about 5.0 cm, about 5.0 mm to about 2.5 cm, about 7.5 mm to about 1.0 cm, or greater than 10 cm, or any value or any intermediate range therebetween. The width of passage or device or both can be, for example, about 5.0 mm, about 6.0 mm, about 7.0 mm, about 8.0 mm, about 9.0 mm, or about 10 mm, or any value or any intermediate range therebetween. In some embodiments, the first channel can be parallel to the second channel and the third channel. Two or more channels in the first channel, the second channel and the third channel can be different from each other in volume, surface area, cross-sectional area, length, width or diameter or any combination thereof. For example, the width, diameter or both of the first channel can be greater than the width, diameter or both of the second channel or the third channel or both. The cross-sectional area, width, diameter or any combination thereof of one or more channels in the first channel, the second channel and the third channel can be different along its length.

[0037] Any channel can contain any number of sections. Any two sections can be continuous or discontinuous. Any two sections can be adjacent or non-adjacent. Any two adjacent sections can be open, closed, partially closed or have a variable connector or any combination thereof. For example, two adjacent sections or two adjacent channels or both can be separated by a valve. Any type of valve can be used. The valve can be a one-way valve or a two-way valve. The channel can be divided into, for example, three sections. For example, one or more channels in the first channel, the second channel, and the third channel can contain a first peripheral section near the first end of the channel, a second peripheral section near the second end of the channel, and a center section between the first peripheral section and the second peripheral section. The channel can have a central longitudinal axis, and each section can have a corresponding central longitudinal axis. These axes can serve as a reference for a relationship, such as the orientation between the corresponding sections of a given channel and the orientation between the sections of different, for example, adjacent channels. The central peripheral section may include a cross-sectional area, width or diameter or any combination thereof greater than one or two peripheral sections in the first peripheral section and the second peripheral section. The central peripheral section of the first channel may include a cross-sectional area, width or diameter or any combination thereof greater than one or two peripheral sections in the first peripheral section and the second peripheral section of the first channel. One or two peripheral sections in the first peripheral section and the second peripheral section of the channel may be colinear with the central section relative to its corresponding central longitudinal axis. One or two peripheral sections in the first peripheral section and the second peripheral section may form an angle other than 180° with respect to its corresponding central longitudinal axis with the central section. For example, the angle may include a right angle, an acute angle or an obtuse angle or any combination thereof. Two or more channels in a plurality of channels may be located in a common plane, for example, the first channel, the second channel and the third channel in a plurality of channels may be located in a common plane.

[0038] The microfluidic device can include any desired number of diaphragms. The number of diaphragms can be, for example, one less than the number of channels. For example, a plurality of channels can include a first channel, a second channel, and a third channel having a first diaphragm and a second diaphragm. Any two diaphragms can vary or be the same in specific parameters, such as geometry, material, or design, or any combination thereof. For example, the length of the first diaphragm can be less than the length of the first channel or the length of the second channel or both. The length of the second diaphragm can be less than the length of the first channel or the length of the third channel or both. The diaphragm can include a film. For example, the first diaphragm, the second diaphragm, or both can include a film. The film can include a porous film. The pore width or diameter or both can be less than about 1.0 nm, about 1.0 nm to about 1.0 cm, about 10 nm to about 1.0 mm, about 10 nm to about 500 μm, about 50 nm to about 100 μm, about 100 nm to about 10 μm, or greater than 1.0 cm or any value or any intermediate range therebetween. The porous membrane may comprise, for example, polydimethylsiloxane (PDMS), polytetrafluoroethylene (PTFE), polyethylene terephthalate (PET), etc. The membrane may comprise a semipermeable membrane. The membrane may comprise a biological membrane or a synthetic membrane or both.

[0039] The first diaphragm may include a first plurality of columns separated by a first plurality of openings, and the second diaphragm may include a second plurality of columns (columns, microcolumns, etc.) separated by a second plurality of second openings, or both. Any desired number, size, shape, and column spacing may be used. For example, the first plurality of columns or the second plurality of columns or both may have a straight cross-section, a curved cross-section, or both. For example, relative to the general shape or the cross-sectional shape or both, the first plurality of columns or the second plurality of columns or both may be circular, elliptical, polygonal, square, triangular, rectangular, pentagonal, hexagonal, heptagonal, octagonal, or trapezoidal, or any combination thereof. The width, diameter, height or depth of post or any combination thereof can be less than 1.0 μm, about 1.0 μm to about 1.0 mm, about 5.0 μm to about 300 μm, about 5.0 μm to about 500 μm, about 5.0 to about 700 μm, about 10 μm to about 250 μm, about 25 μm to about 100 μm, or greater than 1.0 mm or any value or any intermediate range therebetween. Post can be an extension of any surface or combination of surfaces of a channel or adjacent channels. Post can extend from a lateral channel wall, a channel ceiling or a channel floor or any combination thereof. Post can partially or completely span channel width or channel diameter or both. The number, placement, size and shape of posts can affect flow patterns, such as eddy flow, laminar flow and turbulence.

[0040] The columns may be spaced evenly or irregularly to form a plurality of openings between the columns. The average value, for example, the average distance between adjacent columns may be less than 1.0 μm, about 1.0 μm to about 1.0 mm, about 5.0 μm to about 300 μm, about 5.0 to about 500 μm, about 5.0 μm to about 700 μm, about 10 μm to about 250 μm, about 25 μm to about 100 μm, or greater than 1.0 mm, or any value or any intermediate range therebetween. The first plurality of openings may differ in number, size, shape, or spacing, or any combination thereof; or the openings in the second plurality of openings may differ in number, size, shape, or spacing, or any combination thereof; or both. The openings in the first plurality of openings may differ in number, size, shape, or spacing, or any combination thereof, along one or more dimensions of the first diaphragm; or the openings in the second plurality of openings may differ in number, size, shape, or spacing, or any combination thereof, along one or more dimensions of the second diaphragm; or both. The first plurality of openings and the second plurality of openings may differ from each other.

[0041] The device housing may include a substrate, and a plurality of channels and a plurality of channel ports are formed in the substrate. The device housing may include a polymer, a metal or a ceramic or any combination thereof. The polymer may be, for example, silicone. Silicone may include polydimethylsiloxane (PDMS), for example, Campbell et al., "Beyond Polydimethylsiloxane: Alternative Materials for Fabrication of Organ-on-a-Chip Devices and Microphysiological Systems," ACS Biomater. Sci. Eng., 7, 7, 2880-2899 (2021), which is incorporated herein by reference in its entirety. The device housing may be made of polydimethylsiloxane (PDMS), polymethyl methacrylate (PMMA), agarose, alginate, hyaluronic acid, gelatin, other polymer materials, glass or quartz crystal or other optically transparent materials or any combination thereof. The device housing may include a flexible material or a rigid material or both. The device housing may contain conductors or insulators or both.

[0042] Any surface of a microfluidic device or its contents may be coated with one or more materials. The interior, exterior or port of a microfluidic device or any combination thereof may be coated. A substrate by which a microfluidic device may include a first material and one or more channel surfaces may include a second material different from the first material. The second material may be different from the contents of one or more channels in which the second material is present. The second material may be a coating on the first material. One or more channel surfaces may be coated with a variety of different materials. One or more channel surfaces may be coated with a plurality of coatings. At least two of the plurality of coatings may differ in material composition. A substrate may include a first material, and a plurality of coatings may include a first coating adjacent to the substrate on one or more channel surfaces and a second coating covering the first coating, the first coating including a second material, the second coating including a third material. The first material and the second material may be different from each other, and the second material and the third material may be different from each other. The first material, the second material and the third material may be different from each other. A plurality of coatings may include at least two layers having average thicknesses different from each other. A coating may have one, two, three, four, five, or ten or more layers. The coating thickness or patterning or a combination thereof on one or more channel surfaces may differ along the length of one or more channels. Two or more channels in a channel may differ in the material composition of their surface coatings. A first channel may include a first coating, a second channel may include a second coating, and a third channel may include a third coating-the first coating, the second coating, and the third coating differ in material composition, thickness, or patterning or a combination thereof. The thickness of a coating or layer may be one atom, one molecule, less than 1.0 nm, about 1.0 nm to about 1.0 mm, about 10 nm to about 100 nm, about 50 nm to about 5.0 μm, about 0.5 μm to about 50 μm, about 1.0 μm to about 10 μm, about 10 μm to about 100 μm, about 0.5 mm to about 1.0 mm, or greater than 1.0 mm, or any value or any intermediate range therebetween. The patterning can be linear, straight, curvilinear, diagonal, axial, radial, spiral, cross-hatched, spotted, diffuse, random, lateral interior of the channel, bottom interior of the channel, half bottom interior of the channel, top interior of the channel, top half of the channel, top interior of the channel, half top interior of the channel, the first channel, the second channel, and the third channel can be coated with the same material. One or more coatings can contain organic materials or inorganic materials or both. One or more coatings can contain polymers. One or more coatings can contain biomolecules. One or more coatings can contain proteins or nucleic acids, lipids or carbohydrates, or any combination thereof.One or more coatings include fibronectin, an extracellular matrix-derived composition, a basement membrane extract, a cell-free extracellular matrix, a dissolved basement membrane preparation, a mixture of extracellular matrix proteins, a tumor-derived composition, a matrigel or hyaluronic acid or any combination thereof. The substrate may include silicon, and one or more channel surfaces may be coated with a coating comprising silane. One or more channel surfaces may be subjected to silanization. One or more channel surfaces may be treated with plasma. The plasma may include oxygen. One or more coatings may be applied by: emerging, spraying, brushing, injection, precipitation, polymerization or vapor deposition or any combination thereof. One or more channel surfaces may include laminates. One or more channel surfaces may include adhesives.

[0043] One or more surfaces can be etched, shot peened, ground or smoothed or a combination thereof, or otherwise modified to form a pattern, such as a pattern of coatings, ridges, grooves, bumps or holes described herein or other surface features or any combination thereof. Physical surface features can have the size described herein for coatings. One or more coatings can combine one or more physical surface features. Coatings and physical surface features can be hydrophilic or hydrophobic or both. Coatings and physical surface features can affect the flow in one or more channels of a microfluidic device. In addition to or in place of channel surfaces, coatings and physical surface features can be applied to other surfaces of a microfluidic device.

[0044] The device housing may include a transparent material, a translucent material, a tinted material, a colored material, a polarized material, a material that selectively transmits a wavelength of electromagnetic radiation, a material that selectively refracts a wavelength of electromagnetic radiation, a material that selectively reflects a wavelength of electromagnetic radiation, or an opaque material, or any combination thereof. The device housing may include a backing. The backing may include, for example, any material that enables the transmission of light for fluorescence and optical microscopy. The device housing may include a window that allows a view of the interior of the device to be obtained. The view may be a view of one or more channels in a plurality of channels or its components or its contents or any combination thereof. For example, the view may include a view of a first channel or a target object or both. The device housing may include multiple windows that allow multiple views of the interior of the device to be obtained. The microfluidic device may be manufactured using any technology or combination of technologies. For example, the microfluidic device may be manufactured using molding, three-dimensional printing or etching or any combination thereof. Three-dimensional printing may include, for example, selective laser sintering (SLS), fused deposition modeling (FDM) or photopolymerization or any combination thereof.

[0045] The target object can include organic material or inorganic material or both. The target object can include cells, tissues or medical devices or any combination thereof. The cell or tissue can be a human cell. The cell or tissue can be a mammal other than a human, for example, rats, rodents, hamsters, monkeys, apes, dogs, cats, sheep, cattle, horses or pigs. The target object can include spheroids, organoids, biomaterials based on alginate, cell structures supported by scaffolds or bioprinting structures or any combination thereof. Spheroids can be produced using any suitable method, such as, hanging drop method, hydrogel, rotating cell culture, cell suspension with nanofibers, magnetic levitation, microfluidics or bioprinting or any combination thereof. The target object can include a single cell suspension, a suspension of a spheroid or a suspension of an organoid or any combination thereof.

[0046] The target object may comprise a tumor cell, a malignant cell, or a benign cell, or any combination thereof. The target object may comprise a cancer cell or a cancerous tissue, or both. For example, a cancer may be a premalignant growth, a malignant growth, or a tumor caused by abnormal and uncontrolled cell division that may be metastatic or non-metastatic. The cancer can be, for example, breast cancer, prostate cancer, lung cancer, colon cancer, rectal cancer, bladder cancer, non-Hodgkin a lymphoma, melanoma, kidney cancer, pancreatic cancer, oral cancer, pharyngeal cancer, ovarian cancer, thyroid cancer, stomach cancer, brain cancer, glioma, multiple myeloma, esophageal cancer, liver cancer, cervical cancer, laryngeal cancer, intrahepatic bile duct cancer, acute myeloid leukemia, soft tissue cancer, small intestine cancer, testicular cancer, chronic lymphocytic leukemia, Hodgkin lymphoma, chronic myeloid cancer, acute lymphocytic cancer, anal cancer, vulvar cancer, cervical cancer, gallbladder cancer, malignant mesothelioma, bone cancer, joint cancer, hypopharyngeal cancer, eye cancer, nasal cancer, nasal cavity cancer, middle ear cancer, nasopharyngeal cancer, ureteral cancer, peritoneal cancer, omental cancer, mesenteric cancer, or gastrointestinal carcinoid or any combination thereof. The target object can include alginate, alginate derivatives, polyethylene glycol (PEG), poly (lactic acid-co-glycolic acid) (PLGA), poly (ethylene glycol diacrylate) (PEGDA), collagen or hyaluronic acid or any combination thereof. The target object can include, for example, nanoparticles, nanomaterials, polymer biomaterials, ceramic biomaterials, metal biomaterials or surface treatment or any combination thereof. The target object can include a surgical device or a part thereof, such as a suture. The target object can include polypropylene, silk, polyester, polydioxanone, poliglecaprone, polyglactin or nylon or any combination thereof. The target object can be inserted into a microfluidic device using manual techniques, automated techniques or both. For example, pipetting, injection or one or more surgical tools can be used to insert the target object.

[0047] The target may include one or more coatings. The target may include one or more coatings that are different from the substrate. The target may include one or more coatings that are different in material composition, thickness, or patterning, or a combination thereof. The target may be applied with coatings and other surface modifications for channel surfaces described herein, and vice versa. The target may include one or more coatings that are the same as one or more coatings of one or more channel surfaces in terms of material composition, thickness, or patterning, or a combination thereof. The target may be coated with a variety of different materials. The target may be coated with multiple coatings. Multiple coatings may be different in terms of material composition, thickness, or patterning, or a combination thereof. Multiple coatings may include a first coating adjacent to the target and a second coating covering the first coating, the first coating including a first material, and the second coating including a second material. The target may be coated before, during, or after being placed in a microfluidic device. The target coating may be partial or complete. At least 10%, at least 25%, at least 40%, at least 50%, at least 60%, at least 75%, at least 90%, at least 95%, or at least 99%, or any intermediate percentage or any percentage range therebetween, of the target surface may be coated or otherwise modified.

[0048] The matrix can include hydrogels. The matrix can include a mixture of decellularized extracellular matrix, dissolved basement membrane preparations, extracellular matrix proteins, or tumor-derived compositions or any combination thereof. For example, the matrix can include dissolved basement membrane matrix secreted by Engelbreth-Holm-Swarm mouse sarcoma cells; the matrix can include matrigel (MATRIGEL) (Corning). The matrix can include natural polymers, artificial polymers, or cross-linked polymers or any combination thereof. The matrix can include polysaccharides, polypeptides, lipids, polynucleotides, hyaluronic acid, hyaluronic acid methacrylate, hyaluronic acid diacrylate, fibronectin, fibrinogen, collagen, or methacrylate collagen or any combination thereof.

[0049] The component described as being present in a biological composition may also be present in another biological composition or other biological compositions. The first biological composition may include, for example, fibroblasts or endothelial cells or both. Cells that can be used include, for example, B16-F10 cells, MDA-MB-231 cells, MCF-7 cells, HEPG2 cells, etc. or any combination thereof. Cells may include immune cells, RAW264.7 macrophages, L929 fibroblasts, primary cells, dermal fibroblasts, peripheral blood mononuclear cells, pathogenic bacterial cells or non-pathogenic bacterial cells or any combination thereof. Components may include viral particles. Cells may be introduced into microfluidic devices, for example, in the form of cell suspensions in cell culture media, buffers, three-dimensional gels or pre-gel solutions or any combination thereof. Drugs or therapeutic materials (small molecule drugs, macromolecular drugs, antibodies, proteins, nanoparticles or lipids) may be introduced into microfluidic devices, or incorporated into biomaterials or hydrogels placed in microfluidic devices. Introduced cells may include healthy cells or diseased cells or both. The cells may be from a human cell line or a non-human cell line or both. The cells may be from a primary cell culture or an organ explant, etc. The components added to the microfluidic device may include, for example, anti-inflammatory drugs, anti-fibrotic drugs, steroids or immune-related drugs or any combination thereof.

[0050] The first biological composition may include serum, ascorbic acid, retinoic acid, proline, bleomycin, cyclophosphamide, amiodarone, procainamide, penicillamine, gold or nitrofurantoin or any combination thereof. The first biological composition may include compounds, for example, drugs associated with causing fibrosis (e.g., pulmonary fibrosis or liver fibrosis or both). The first biological composition may include environmental substances, for example, asbestos, silicon dioxide, titanium dioxide, zirconium oxide, graphite, coal dust, mineral dust or any combination thereof. The first biological composition may include bacteria, fungi, components thereof or combinations thereof. The first biological composition may include biomolecules, complement proteins, coagulation-related proteins, bacterial cell wall proteins or lipopolysaccharides (LPS) or any combination thereof that promote fibrosis. The second biological composition may include at least one type of biological cell that can promote fibrosis. The second biological composition can include factors that promote biological cell differentiation into biological cell types that can promote fibrosis, factors that promote the expression of biological molecules that promote fibrosis, or factors or any combination thereof that promote the amplification of biological cell types that can promote fibrosis. The second biological composition can include serum, platelets, leukocytes, macrophages, monocytes, peripheral blood mononuclear cells (PBMC), stem cells, mast cells, dendritic cells, neutrophils, granulocytes, T cells or B cells or any combination thereof. The third biological composition can include molecular populations that can form gradients that are radiated from the third channel to the first channel or to the first channel and the second channel. The third biological composition can include cytokines, chemokines, cell nutrients, transcription factors or growth factors or any combination thereof. The third biological composition can include, for example, CCL-2 / MCP-1, CCL3, CCL11, fibroblast growth factor (FGF), interferon, IFN-γ, interleukin (IL), IL-4, IL-1, IL-2, IL-5, IL-6, IL-7, IL-9, IL-13, IL-17A, IL-17F, IL-21, IL-22, IL-25, platelet-derived growth factor (PDGF), transforming growth factor (TGF)-α, transforming growth factor (TGF)-β, granulocyte colony stimulating factor (G-CSF), macrophage colony stimulating growth factor (M-CSF), granulocyte macrophage colony stimulating factor (GM-CSF), prothrombin or thrombin or any combination thereof. At least one type of molecule of the gradient can include cytokines, chemokines, cell nutrients, transcription factors or growth factors or any combination thereof. At least one type of molecule of the gradient can include transforming growth factor beta (TGF-β). TGF-β may comprise TGF-β1, TGF-β2 or TGF-β3 or any combination thereof.The first biological composition, the second biological composition or the third biological composition or any combination thereof may comprise labeled cells or labeled biomolecules or both. The first biological composition, the second biological composition or the third biological composition or any combination thereof may comprise a compound with known or potential anti-fibrotic efficacy.

[0051] The fibrosis diagnostic system can be used for modeling one or more fibrosis.Fibrosis can be associated with any tissue or organ, such as lung, heart, liver, pancreas, kidney, bladder, skin, skeletal muscle or smooth muscle or any combination thereof.The example of fibrosis can include endomyocardial fibrosis, cystic pancreatic fibrosis, diatomaceous earth fibrosis, diffuse interstitial fibrosis, idiopathic fibrosis, endomyocardial fibrosis, graphite fibrosis, idiopathic pulmonary fibrosis, idiopathic retroperitoneal fibrosis, mediastinal fibrosis, neoplastic fibrosis, nodular subepidermal fibrosis, full wall bladder fibrosis, periureteral fibrosis, fibrin formation after fibrosis, hyperplastic fibrosis, pulmonary fibrosis, alternative fibrosis, retroperitoneal fibrosis or nerve root cuff sample fibrosis or any combination thereof.Fibrosis can be the result of tissue autotransplantation, tissue allograft or tissue xenotransplantation or any combination thereof.Fibrosis can be the result of inorganic material (e.g., metal, ceramic, mineral or any combination thereof). The feature of fibrosis can be the deposition of collagen on target object.One or more types of collagen can characterize fibrosis.Collagen can comprise for example, collagen I, collagen II, collagen III, collagen IV, collagen V, collagen VI, collagen, collagen VII, collagen VIII, collagen IX, collagen X, collagen XI, collagen XII, collagen XIII, collagen XIV, collagen XV, collagen XVI, collagen XVII, collagen XVIII, collagen XIX, collagen XX, collagen XXI, collagen XXII, collagen XXIII, collagen XXIV, collagen XXV, collagen XXVI, collagen XXVII or collagen XXVIII or any combination thereof.One or more types of procollagen or collagen metabolites or both can characterize fibrosis.

[0052] The fibrosis diagnosis system can be preloaded with one or more components, such as reagents or target objects or both. For example, the fibrosis diagnosis system can include a matrix, a first biological composition, a second biological composition or a third biological composition or any combination thereof. The fibrosis diagnosis system can include a target object. The fibrosis diagnosis system can include a gradient of at least one molecule across the first channel, the second channel or the third channel or any combination thereof. One or more components of the matrix, the first biological composition, the second biological composition or the third biological composition or any combination thereof are anchored to the channel surface.

[0053] The fibrosis diagnostic system may include a detector. The detector may include a sensor. The sensor may be configured to detect fibrosis of a target object or indicate characteristics of fibrosis of a target object. The detector may be configured to receive a microfluidic device, for example, in a receiving portion or on a stage of a microscope. The detector may include a detector housing, which includes the interior of the detector. The detector may include a receiving portion configured to receive the microfluidic device. For example, the receiving portion may include an opening in the detector housing. Any number or any type or both of receiving portions may be used. The receiving portion may be configured to receive one or more microfluidic devices. Multiple receiving portions may be configured to receive corresponding multiple microfluidic devices.

[0054] The fibrosis diagnostic system may include a thermal subsystem. The thermal subsystem may include a heater or a cooler or both. The thermal subsystem may be configured to adjust the temperature of the microfluidic device. The detector may include a thermal subsystem. The thermal subsystem may be configured to, for example, adjust the temperature of the microfluidic device when the microfluidic device is in a receiving portion.

[0055] The fibrosis diagnosis system may include a fluid subsystem configured to be connected to a microfluidic device fluid. The fluid subsystem may be configured to load one or more components into the microfluidic device. The fluid subsystem may be configured to supply solid, liquid or gas or any combination thereof to the microfluidic device. The fluid subsystem may be configured to load a target object, a matrix, a first biological composition, a second biological composition or a third biological composition or any combination thereof into the microfluidic device. The fluid subsystem may be configured to supply one or more nutrients to the microfluidic device. The fluid subsystem may be configured to, for example, regulate the pressure in the internal microfluidic device in one or more channels in the channel. The pressure of the inside (e.g., channel) of the microfluidic device may be the same as atmospheric pressure, higher than or lower than atmospheric pressure. A pressure difference may be applied between channels, in the channel or both. A pressure equal to, higher than or lower than the osmotic pressure may be applied. The fluid subsystem may be configured to regulate the gradient across the first channel, the second channel or the third channel or any combination thereof. The gradient may be controlled, for example, by adding gradient solutes / molecules or applying pressure or both. A pressure equal to, higher than or lower than the osmotic pressure associated with the gradient molecule may be applied. The fluid subsystem can be configured to remove one or more components from the microfluidic device. The detector can, for example, include a fluid subsystem. The fluid subsystem can be configured to be, for example, communicated with the microfluidic device fluid when the microfluidic device is in the receiving portion. The fluid subsystem can include a channel, a valve, a manifold or a pump or any combination thereof. The fluid subsystem can include a handheld pipette. The fluid subsystem can be configured to provide a desired flow rate of a solid, a liquid or a gas or any combination thereof through one or more channels in a plurality of channels. The flow rate can be, for example, less than about 0.001 microliter / second, about 0.001 microliter / second to about 100 milliliters / second, about 0.01 microliter / second to about 10 milliliters / second, about 0.1 microliter / second to about 1.0 milliliters / second, about 1.0 microliter / second to about 100 microliters / second, about 10 microliters / second to about 50 microliters / second, or greater than 100 milliliters / second, or any value or any intermediate range therebetween. The fluidic subsystem may contain or be configured to be fluidically connected to a supply of one or more reagents, such as multiple reservoirs or a single reservoir.

[0056] The sensor may be configured to detect particles or waves or both indicating fibrosis emitted from the target object or the microfluidic device or both. The sensor may be configured to detect electromagnetic radiation. The electromagnetic radiation may include, for example, ultraviolet radiation, visible light or infrared radiation or any combination thereof. The electromagnetic radiation may include fluorescence or phosphorescence or both. The detector may include a particle source or a wave source or both, the particle source or the wave source or both being configured to transmit particles, waves or both to the target object or the microfluidic device or both. The fibrosis diagnostic system may include an electromagnetic radiation source. For example, the detector may include an electromagnetic radiation source. The electromagnetic radiation source may include, for example, a light emitting diode, a laser, an incandescent light source, an arc light source, a halogen light source, a chemical vapor light source or a fluorescent light source or any combination thereof. The detector may include an optical subsystem, the optical subsystem being configured to transmit radiation or affect radiation or both. The optical subsystem may include one or more lenses, one or more filters, one or more mirrors or one or more diffraction gratings or any combination thereof. The optical subsystem or the sensor or both may include a microscope or one or more components thereof. The optical subsystem may include a sensor or an electromagnetic radiation source or both. The sensor may include a microphone, and the fibrosis diagnostic system may include a source of sound waves. The sound waves may include ultrasound, and the microphone may detect ultrasound. The sensor may include a force sensor, an actuator, or both. Fibrosis of the target object may result in a fibrotic structure having a tensile strength or a compressive strength, or both, that can be detected by the sensor.

[0057] The sensor may be configured to generate an output signal based on an input signal detected by the sensor. The detector may include a transmitter, a receiver, a transceiver, or any combination thereof, wherein the transmitter, the receiver, the transceiver, or any combination thereof is configured to receive information or send information or both. The information may include an output signal. The information may include an operation signal for operating a sensor or other component of the system. The information may be transmitted through a network. The network may be a local network or a long-distance network. The network may be wired or wireless or both. The detector or other component of the system may include a data port. Any desired number of data ports may be provided. The system may further include a processor configured to receive and process information. The processor may be configured to control one or more subsystems or components of the fibrosis diagnosis system. For example, the processor may be configured to control a sensor, an electromagnetic radiation source, an optical subsystem, a thermal subsystem, or a fluid subsystem, or any combination thereof. The processor may be configured to receive an output signal from a sensor, receive an operation signal from a user input, or both. The detector may include a processor, for example. The processor may include a local processor or a remote processor, or both. The processor may be a microprocessor. The processor may be a central processing unit (CPU). The processor may include one or more processing chips. The system may include a memory configured to communicate with the processor. The memory may be local or remote. The memory may be temporary or non-temporary or both. The memory may include random access memory (RAM) or read-only memory (ROM) or both. The memory may be configured to store programs that control the fibrosis diagnosis system or its components. The processor may, for example, include the memory. The detector may, for example, include the memory.

[0058] The fibrosis diagnosis system may include a user interface. The system may include any number of user interfaces or any kind of user interface or both. The user interface may be interactive or non-interactive or both, for example, the user interface may include one or more components that are interactive and one or more components that are not interactive. The user interface may be stand-alone or integrated into one or more system subsystems or components. The user interface may include a monitor, a button, a keyboard, a mouse, a light signal, a camera, an eyepiece lens, a speaker or a microphone or any combination thereof. The monitor may include, for example, a touch screen. The camera may be a still camera or a video camera or both. The eyepiece lens may be part of a microscope system or subsystem. Two eyepiece lenses may be provided, for example to allow binocular viewing. The detector may, for example, include a user interface.

[0059] The fibrosis diagnostic system may include a power source or a power interface or both. The power source may include a battery, such as a rechargeable battery. For example, the power interface may include an inverter for receiving an alternating current wall current and outputting a direct current. The detector may include a power source or a power interface or both, for example.

[0060] The fibrosis diagnosis system may include a kit or be provided as a kit. The kit may include two or more components of the system. The kit may include the supply of multiple specific system components (e.g., multiple multifluidic devices) in a specific system component. The kit may include, for example, a microfluidic device and one or more reagents for use in a microfluidic device. The kit may include, for example, a microfluidic device; and a matrix, a first biological composition, a second biological composition, or a third biological composition, or any combination thereof. One or more components or subsystems of the fibrosis diagnosis system, whether or not supplied as a kit, may include an identifier. The identifier may include a serial number, a product number, a barcode, an RFID tag, a color code, a name, a digital or alphanumeric code, or any combination thereof.

[0061] The present disclosure provides methods for using a fibrosis diagnostic system, its subsystems, and its components. For example, a method for analyzing the susceptibility of a target object to fibrosis is provided. The method may include one or more of the following steps. The target object, a matrix, and a first biological composition may be placed in a first channel of a microfluidic device. A second biological composition may be placed in a second channel of the microfluidic device. A third biological composition may be placed in a third channel of the microfluidic device. The target object or the first channel or both may be analyzed to determine the presence or absence of fibrosis associated with the target object.

[0062] The analysis can be carried out after the predetermined time period after the target object, matrix, the first biocomposition, the second biocomposition or the third biocomposition or any combination thereof is placed in the microfluidic device. The predetermined time period can be enough to allow the gradient of at least one type of molecule to be formed. The analysis can be carried out twice or more. Any two continuous analyses can be separated by a predetermined time period. The time period can be counted as seconds, minutes, hours, days or weeks. The time period can be, for example, less than about one minute, about one minute to about one month, about one hour to about two weeks, about three hours to about one week, about six hours to about three days, about 12 hours to about one day, greater than about one month or any time period therebetween or the time period range therein. The analysis can comprise removing one or more components from the microfluidic device.

[0063] The analysis may comprise an immunoassay. The analysis may comprise the use of an antibody. The antibody may be labeled with a detectable label. The analysis may comprise applying a primary antibody and a secondary antibody comprising a detectable label and bound to the primary antibody. The primary antibody may be bound to collagen, α-smooth muscle actin, or both. The analysis may comprise a histological assay. Cells and / or tissues may be fixed, for example, prior to staining, for assay. Any suitable fixative may be used, for example, formaldehyde fixative, neutral buffered formalin (NBF), paraformaldehyde (PFA), or methanol. The fixative may be an aldehyde fixative, a mercury fixative, an alcohol fixative, an oxidant fixative, or a picrate fixative, or any combination thereof. Additional examples of fixatives are acetone, phosphate buffered formalin, zinc formalin, calcium methylal, saline methylal, alcohol formalin, alcohol methylal acetate, zinc formalin, Alcian Blue, Bouin's solution, B-5 ​​fixative, Camoy's solution, Clarke's solution, Gendre's solution, Helly's fixative, Hollande's solution, methacarn and Zenker's fixative. Cells and / or tissues may be partially or completely dehydrated for determination. Analysis may include embedding or antigen retrieval or both. Any suitable stain or combination of stains may be used. The stain may include a dye. Picric acid may be used. Analysis may include, for example, application of Sirius red dye alone or in combination with one or more additional stains. Examples of stains include carmine, hematoxylin, hematoxylin, fluorescein, eosin, silver nitrate, Giemsa stain, Golgi stain, Gram stain, Kluver-Barrera stain, Mallory's CT stain, Masson's stain, periodic acid-Schiff (PAS) stain, Romanowsky stain, trichrome stain, AZAN trichrome stain, toluidine blue, and Verhoeff's Van Gieson stain, and any combination thereof.

[0064] The analysis may include visually inspecting the target object or the first channel or both for one or more indications of fibrosis associated with the target object. The analysis may include the use of microscopy. The analysis may include the use of one or more spectroscopic techniques. For example, the analysis may include nonlinear optical (NLO) multiphoton microscopy, second harmonic generation (SHG) imaging, third harmonic generation (THG) imaging, two-photon excitation fluorescence (TPEF), two-photon excitation fluorescence (TPEF), cell tracker dye imaging, visible light imaging, (ELISA), Griess assay, histopathology, immunostaining, cleared tissue microscopy, or proteomic profiling of a homogenized hydrogel, or any combination thereof. The analysis may include the use of a detector to detect one or more indications of fibrosis associated with the target object. The analysis may include inserting a microfluidic device into a receiving portion.

[0065] The method may include preparing a target object, a matrix, a first biological composition, a second biological composition or a third biological composition or any combination thereof from a patient sample. The method may include implanting the target object into a patient or a patient. For example, the method may include implanting a medical device or a tissue or both into a patient or a patient, wherein the target object is a representative model of the medical device or the tissue or both. The method may include removing a medical device or a tissue or both from a patient, wherein the target object is a representative model of the medical device or the tissue or both.

[0066] The method may include administering an antifibrotic agent to a patient. The analysis may include determining a quantitative measure of fibrosis. The dosage of the antifibrotic agent administered to a patient may be based on a quantitative measure of fibrosis. The antifibrotic agent is administered to a patient before, during, or after implantation of a target object, a tissue associated with the target object, or a medical device associated with the target object, or any combination thereof. In addition to or in lieu of the antifibrotic agent, one or more therapeutic agents may be administered to the patient. The patient may be, for example, a mammalian patient or a human patient or both.

[0067] A method for screening a therapy for efficacy in treating fibrosis is provided. The method may comprise one or more of the following steps. A target object, a matrix, and a first biological composition may be placed in a first channel of a microfluidic device. A second biological composition may be placed in a second channel of the microfluidic device. A third biological composition may be placed in a third channel of the microfluidic device. The therapy may be performed in the microfluidic device. The target object or the first channel or both may be analyzed to determine the presence, absence, or extent of fibrosis associated with the target object. The efficacy of the therapy may be determined based on a difference in the presence, absence, or extent of fibrosis without the therapy. The therapy may comprise a therapeutic compound, and performing the therapy comprises placing the therapeutic compound in the first channel, the second channel, or the third channel, or any combination thereof. The therapeutic compound may comprise a pharmaceutical compound or a biologic, or both. The therapy may comprise modification of the target object. The therapy housing comprises placing a medical device in the first channel, the second channel, or the third channel, or any combination thereof. The therapy may comprise applying radiation to the microfluidic device. The method may comprise treating a patient with the screened therapy. The method of screening provided herein comprises a high throughput screening method.

[0068] The present disclosure provides a method for screening therapy for the efficacy of treating cancer. The method may include one or more of the following steps. A target object, a matrix and a first biological composition may be placed in a first channel of a microfluidic device, wherein the target object may include tumor cells. A second biological composition may be placed in a second channel of a microfluidic device. A third biological composition may be placed in a third channel of a microfluidic device. The therapy may be performed on a microfluidic device. The target object or the first channel or both may be analyzed to determine the presence, absence or degree of fibrosis associated with the target object. The efficacy of the therapy may be determined based on the presence, absence or degree of cancer without therapy. Other screening methods using a fibrosis diagnostic system are also provided. For example, the screening method may be used to monitor immune phenomena commonly associated with foreign matter and more specific phenomena, such as graft versus host disease (GVHD) or autoimmune syndrome.

[0069] A non-transitory computer-readable medium is provided by the present disclosure. For example, a non-transitory computer-readable medium storing a program is provided, the program comprising instructions for performing the method of the present disclosure. A non-transitory computer-readable medium storing a program is provided, the program comprising instructions for controlling a fibrosis diagnostic system or subsystem or a component thereof or any combination thereof.

[0070] Figure 1A1 is a perspective view of a microfluidic device 110 according to the present disclosure. The microfluidic device 110 can be used in the fibrosis diagnosis system or method or both of the present disclosure. The microfluidic device 110 can include a device housing 112, an outer surface 114 of the device housing 112. The device housing 112 can define a device interior 116 that can include multiple channels 118. Multiple channels 118 can include a first channel 120 between a second channel 140 and a third channel 160. The first diaphragm 130 can separate the first channel 120 from the second channel 140 or otherwise demarcate. The second diaphragm 150 can separate the first channel 120 from the third channel 150 or otherwise demarcate. The first diaphragm 130 and the second diaphragm 150 can have any design, for example, designs such as porous membranes or column (microcolumn) arrays described herein. The first channel 120 can include an interior 122, a first end 123, a first channel port 124 near the first end 123, a second end 125, and a second channel port 126 near the second end 125. The first channel 120 can include a surface 127, and the surface 127 can be modified, for example, with a surface modification 129, which is configured to receive and retain one or more target objects. The first channel 120 can include a third channel port 118 between the first channel port 124 and the second channel port 126 to facilitate insertion and retrieval of one or more target objects from the interior 122. The second channel 140 can include an interior 142, a first end 143, a first channel port 144 near the first end 143, a second end 145, and a second channel port 146 near the second end 145. The third channel 160 can include an interior 162, a first end 163, a first channel port 164 near the first end 163, a second end 165, and a second channel port 166 near the second end 165. The channel ports can provide a pathway between the interior of the channel, through the device housing 112 , to the device surface 116 , and areas external to the microfluidic device 110 . Figure 1B yes Figure 1A A plan view of the microfluidic device shown in FIG.

[0071] Figure 2Ais a perspective view of a fibrosis diagnostic system 200 showing a microfluidic device 210 detached from a detector 270 according to the present disclosure. The microfluidic device 210 may include features such as those described for the microfluidic device 110 or any other microfluidic device described herein. The detector 270 may include a detector housing 272, a detector surface 274, a detector interior 276, and a receptacle 278 that allows all or part of the microfluidic device 210 to enter the detector interior 276. The depiction of the detector 270 is for discussion purposes, and other designs suitable for a given embodiment may be employed. For example, the detector 270 may employ a more open design, such as a design for a microscope having an open receptacle in the form of a stage for placing the microfluidic device 210. The detector 270 may include a user interface 280, and as Figure 2A , 2B Shown is for illustrative purposes and is non-restrictive.The user interface of system 200 can be integrated into detector 270, or provided as a stand-alone component or both.User interface 280 is shown as including elements such as monitor 282, button 284, signal 286,287 and data port 288,289.Screen 282 can include, for example, a screen.The screen can be provided as a touch screen.The screen can display a graphical user interface (GUI).Button 284 can, for example, allow one or more functions of control detector 270, for example, to connect / disconnect detector 270 or to start imaging of microfluidic device 210 or both.Single button is shown, but any number of buttons can be provided or omitted, for example, buttons for corresponding functions or multiple buttons in the form of keyboard or keypad.One or more buttons can be replaced with other types of actuators (such as toggle switches).Signals 286,287 can include light, for example, light of corresponding colors. The corresponding signal can correspond to a specific operating state of, for example, the detector 270, such as a power state, the engagement of the microfluidic device 210, or the imaging of the microfluidic device 210, or any combination thereof. Alternatively or in addition, the signal can change the color corresponding to the specific operating state. Two signals are shown, but any number of buttons can be provided or omitted. One or more signals can take the form of a speaker or a microphone or both. Data ports 288, 289 can provide for any number of different functions, such as data transmission, power transmission, and connection to peripheral user interface components. Two data ports are shown, but any number of data ports can be provided or omitted. In an embodiment comprising a microscope, the user interface 280 can include one or more eyepieces configured to view the microfluidic device 210 for the user.

[0072] Figure 2ASeveral illustrative breaks away from the detector housing 272 are included to illustrate various components that may be located within the detector interior 276. For example, a fluid subsystem 279 is shown, which can be configured to supply various components to the microfluidic device 210 while being held in the receiving portion 278. A thermal subsystem 299, such as a heater, is shown, which is configured to regulate the temperature of the microfluidic device 210 while being held in the receiving portion 278. A sensor 290 is shown, which can detect one or more signals from the microfluidic device 210, such as emitted radiation generated by excitation radiation emitted by a radiation source 291. The number and placement of sensors and light sources can vary. An optical subsystem 292 is also shown, and the optical subsystem can include one or more optical elements, such as lenses, mirrors, diffraction gratings, and filters. The optical subsystem 292 can include one or both of the sensor 290 and the radiation source 291. A processor 294 is shown, which can process information related to the operation of the detector 270. A transceiver 296 is shown which can receive and transmit signals to or from the user interface 280, sensor 290, processor 294, a network, for example, via data ports 288, 289 or a wireless network, or any combination thereof. A power supply 298 is shown which can take the form of a stand-alone power supply or a power supply interface for transmitting / converting power from an external power source, or both. Figure 2B FIG. 2 is a diagram showing a microfluidic device 210 coupled to a detector 270 via a receiving portion 278 according to the present disclosure. Figure 2A A perspective view of a fibrosis diagnostic system 200 is shown.

[0073] Figure 3 310 according to the present disclosure. The microfluidic device 310 can be used in the fibrosis diagnostic system or method or both of the present disclosure. The microfluidic device 310 can include a device housing 312, an outer surface 314 of the device housing 112. The device housing 312 can define a device interior 316 that can include multiple channels. The multiple channels can include a first channel 320 between a second channel 340 and a third channel 360. The first diaphragm 330 can separate or otherwise demarcate the first channel 320 from the second channel 340. The first diaphragm 330 is shown as having a microcolumn design, the microcolumn design including multiple columns (microcolumns) 332 defining multiple openings 334 therebetween. The second diaphragm 350 can separate or otherwise demarcate the first channel 320 from the third channel 350. The second diaphragm 350 is shown as having a microcolumn design, the microcolumn design including multiple columns (microcolumns) 352 defining multiple openings 354 therebetween. The multiple channels can include and Figure 1A118 in the microfluidic device 310. For example, the first channel 320 can include a first channel port 324 near the first end, a second channel port 326 near the second end, and a third channel port 318 between the first channel port 324 and the second channel port 326 to facilitate the insertion and retrieval of one or more target objects from a container 329 in the first channel 320. The second channel 340 can include a first channel port 344 near the first end and a second channel port 346 near the second end. The third channel 360 can include a first channel port 364 near the first end and a second channel port 366 near the second end. The channel ports can provide a passage between the interior of the channel through the device housing 312 to the device surface 316 and the area outside the microfluidic device 310.

[0074] Figure 4 4 is a plan view of a microfluidic device 410 according to the present disclosure. The microfluidic device 410 can be used in the fibrosis diagnosis system or method or both of the present disclosure. The microfluidic device 410 can include a device housing 412, an outer surface 414 of the device housing 412. The device housing 412 can define a device interior 416 that can include multiple channels. The multiple channels can include a first channel 420 between a second channel 440 and a third channel 460. The first diaphragm 430 can separate or otherwise demarcate the first channel 420 from the second channel 440. The second diaphragm 450 can separate or otherwise demarcate the first channel 420 from the third channel 450. The multiple channels can include Figure 1A118 in the plurality of channels 118. Further, one or more channels in the channel can be divided into channel sections, wherein the corresponding sections are different from each other in one or more parameters. For example, the first channel 420 can include a first channel port 424 near the first end, a second channel port 326 near the second end, and a third channel port 418 between the first channel port 424 and the second channel port 426 to facilitate the insertion and retrieval of one or more target objects from the container 429 in the first channel 420. The first channel 420 can have three sections 420a, 420b and 420c. The first channel port 424 can be located in the second section 420b, the second channel port 426 can be located in the third section 420c, and the third channel port can be located in the first section 420a. The first section 420a can be adjacent to both the diaphragms 430 and 450 along its length. The second channel 440 can include a first channel port 444 near the first end and a second channel port 446 near the second end. The second channel 440 may have three sections 440a, 440b, and 440c, wherein the first channel port 444 is located in the second section 440b and the second channel port 446 is located in the third section 440c. The second section 440b may have an angle α relative to the first section 440a. The third section 440c may have an angle β relative to the first section 440a. Angles α and β may be the same or different. The angles may be acute, right, or obtuse. The second section 440b may have an angle psi relative to the second section 420b. The third section 440c may have an angle chi relative to the third section 420c. Angles psi and chi may be the same or different. The angles may be acute, right, or obtuse. The third channel 460 may include a first channel port 464 near the first end and a second channel port 466 near the second end. The third channel 440 can have three sections 460a, 460b and 460c, wherein the first channel port 464 is located in the second section 460b and the second channel port 466 is located in the third section 460c. The second section 460b can have an angle γ relative to the first section 460a. The third section 460c can have an angle δ relative to the first section 460a. Angles γ and δ can be the same or different. The angle can be the same or different from angles α and β. The angle can be an acute angle, a right angle, or an obtuse angle. The second section 460b can have an angle phi relative to the second section 420b. The third section 460c can have an angle ω relative to the third section 420c. Angles phi and ω can be the same or different. The angle can be the same or different from psi and chi. The angle can be an acute angle, a right angle, or an obtuse angle. The channel port can provide a passage between the area outside the microfluidic device 410 and the device surface 416 through the device housing 412 inside the channel.

[0075] Figure 5 5 is a plan view of a microfluidic device 510 according to the present disclosure. The microfluidic device 510 can be used in the fibrosis diagnosis system or method or both of the present disclosure. The microfluidic device 510 can include a device housing 512, an outer surface 514 of the device housing 512. The device housing 512 can define a device interior 516 that can include multiple channels. The multiple channels can include a first channel 520 between a second channel 540 and a third channel 560. The first diaphragm 530 can separate or otherwise demarcate the first channel 520 from the second channel 540. The second diaphragm 550 can separate or otherwise demarcate the first channel 520 from the third channel 550. The multiple channels can include Figure 1A118 in the plurality of channels 118. Further, one or more channels in the channel can be divided into channel sections, wherein the corresponding sections are different from each other in one or more parameters. For example, the first channel 520 can include a first channel port 424 near the first end, a second channel port 526 near the second end, and a third channel port 518 between the first channel port 524 and the second channel port 526 to facilitate the insertion and retrieval of one or more target objects from the container 529 in the first channel 520. The first channel 520 can have three sections 520a, 520b and 520c. The first section 520a can include three regions 520a1, 520a2 and 520a3. The first channel port 524 can be located in the second section 520b, the second channel port 526 can be located in the third section 520c, and the third channel port can be located in the first section 520a. The first region 520a1 can be adjacent to both the diaphragms 530 and 550 along its length. Region 520a2 may be between region 520a1 and second section 520b. Region 520a3 may be between region 520a1 and third section 520c. Regions 520a2 and 520a3 may vary in cross-sectional area, width or diameter or any combination thereof along their respective lengths. For example, regions 520a2 and 520a3 may be defined by angles θ and rho, respectively. Angles θ and rho may be the same or different. The angles may be acute, right or obtuse. Second channel 540 may include a first channel port 544 near the first end and a second channel port 546 near the second end. Second channel 540 may have three sections 540a, 540b and 540c, wherein the first channel port 544 is located in the second section 440b, and the second channel port 546 is located in the third section 540c. The third channel 560 may include a first channel port 564 near the first end and a second channel port 566 near the second end. The third channel 540 can have three sections 560a, 560b, and 560c, with a first channel port 564 located in the second section 560b and a second channel port 566 located in the third section 560c. The channel ports can provide access between the interior of the channel through the device housing 512 to the device surface 516 and areas outside the microfluidic device 510.

[0076] Fig. 6A is a top perspective view of a microfluidic device 610 according to the present disclosure. Figure 6B yes Fig. 6A 610 is a front perspective view of the microfluidic device 610 shown in FIG. Figure 6C yes Fig. 6A An enlarged perspective view of a portion of a microfluidic device 610 is shown in FIG. Fig.6D yes Fig. 6AAn enlarged plan view of a portion of a microfluidic device 610 is shown in FIG. Fig. 6E yes Fig.6D A further enlarged plan view of a portion of the enlarged view shown in . Figures 6A-6E This is discussed in detail in Example 1 below. Fig. 7A is a schematic cross-sectional view of a loaded microfluidic device 710 at a first point in time in accordance with the present disclosure. Figure 7B At the second time point Fig. 7A Schematic cross-sectional view of a loaded microfluidic device 710 is shown in FIG. Fig. 7A and 7B This is discussed in detail in Example 1 below.

[0077] Figure 8 800 is a schematic diagram of a fibrosis diagnosis system 800 according to the present disclosure. The fibrosis diagnosis system 800 may include various components and subsystems, such as a microfluidic device 810, a reagent 820, a target object 830, a fluid subsystem 840, a thermal subsystem 850, a sensor 860, a processor 870, a memory 880, and a user interface 890. These various components and subsystems may be arranged, configured, combined, multiplied, or omitted or any combination thereof without departing from the present disclosure. One or more components or subsystems of the system may be disposed in one or more sterile enclosures or other enclosures that separate their contents from the surrounding environment. Fig. 9 and 10 Two examples of such systems are depicted.

[0078] Fig. 9 is a schematic diagram of a fibrosis diagnosis system 900 according to the present disclosure. In the system 900, a reagent kit 905 is provided, the reagent kit comprising a microfluidic device 910 and a reagent 920. A target object preparation subsystem 915 is provided, the target object preparation subsystem is configured to form a target object 930 for insertion into the microfluidic device 910 together with the reagent 920. A microfluidic device loader 925 is provided, the microfluidic device loader may include a microfluidic subsystem for loading the reagent 920 or the target object 930 or both into the microfluidic device 910. An incubator 935 is provided, the incubator may include a thermal subsystem 950, the thermal subsystem is configured to regulate the temperature of the microfluidic device 910 and its contents during the fibrosis diagnosis assay. A detector 945 comprising a sensor 960 is provided, the detector is used to image the microfluidic device 910 or its contents or both during the fibrosis diagnosis assay. A computing device 945 is provided that includes a processor 970 , a memory 980 , and a user interface 990 that can be used to control the system 900 and process information related thereto. One or more system components or subsystems can be interconnected via a network 955 .

[0079] Fig.10 is a schematic diagram of a fibrosis diagnostic system 1000 according to the present disclosure. The system may include a loaded microfluidic device 1065, which includes a microfluidic device 1010 containing a reagent 1020 and a target object 1030. The system may include a detector 1075, which includes a fluid subsystem 1040, a thermal subsystem 1050, a sensor 1060, a processor 1070, a memory 1080, and a user interface 1075. After a period of time has passed since the loaded microfluidic device 1010, the loaded microfluidic device 1065 or a component thereof may be inserted into or onto the detector 1075 to analyze the target object 1030 for possible fibrosis.

[0080] Example 1

[0081] A three-channel microfluidic device 610, 710 ( Figures 6A-6E , 7A and 7B) for co-culture of fibroblasts and macrophages with target objects (721) embedded in a matrix (722a), such as three-dimensional cancer spheroids or biomaterials. Microfluidic molds / housings 612, 712 are prepared using standard photolithography techniques, and the device is made of polydimethylsiloxane (PDMS). The central (first) channel 620 contains a three-dimensional matrix gel embedded with fibroblasts, and the biomaterial implant or three-dimensional spheroid is placed in the three-dimensional matrix gel embedded with fibroblasts through holes (ports) 628, 728. The outer (second and third) channels 640, 660 (740, 760) are used to introduce macrophages and chemokines, respectively. Each channel is connected by a corresponding diaphragm 630, 650 (730, 750), a micropillar array 652 (in Figure 6B As can be seen in FIG. 6 , the corresponding microcolumn 632 is Fig.6D and 6E), 752 are separated from adjacent channels, thereby forming multiple openings 654 (634) therebetween. The recesses 629, 729 below the central channels 620, 720 enable biomaterials or spheroids to be safely placed in the device. The general device design allows the introduction of a combination of multiple cell types, three-dimensional, two-dimensional and fluid cultures, as well as the introduction of various biomaterials, nanomaterials, cytokines / chemokines, therapeutic drugs and other stimulants that may affect fibrosis formation. The device design can also be modified by adjusting the number of channels, channel height, channel length, channel width, notch height, notch shape, type of microcolumns, size of microcolumns, spacing between adjacent microcolumns and other aspects of the device feature size. Using fluorescently labeled molecules of different sizes, a cytokine gradient can be generated throughout the device 610, thereby enabling cells to be potentially stimulated with cytokines. Device 610 and similar devices can be used to study the fibrosis of various target objects (e.g., melanoma spheroids and alginate-based biomaterials).

[0082] Example 2

[0083] L929 mouse fibroblasts and RAW264.7 mouse macrophages are colored red and green, respectively. (Thermo Fisher Scientific) dye fluorescent labeling. Extensive studies were conducted to optimize cell seeding density, culture time, and biomaterial implantation. The cells were co-cultured in two dimensions or co-cultured in the microfluidic device described in Example 1, after which cancer spheroids or alginate-based biomaterials were implanted in two-dimensional cultures and microfluidic devices. Immunostaining, fluorescence microscopy, Gliese assay, and histological analysis were used to evaluate immune cell activation and fibrosis formation in response to spheroids and implants. Gliese assays were performed on culture media collected from cultured cells. Co-culture of RAW264.7 macrophages with different ratios of L929 fibroblasts resulted in similar levels of nitrite production as culture of RAW264.7 macrophages alone, indicating that L929 cells do not activate RAW264.7 cells. In contrast, stimulation of macrophages with lipopolysaccharide (LPS) resulted in macrophage activation, as indicated by nitrite production. With Dye visualization of co-culture of RAW264.7 and L929 cells. No morphological changes were observed due to co-culture. F-actin visualization was performed on RAW264.7 cells treated with LPS. Morphological changes were observed with increasing LPS concentrations. Co-culture of L929 and RAW264.7 cells was performed in a microfluidic device. L929 cells were cultured in 3D within a Matrigel channel, while macrophages were introduced into 2D from an adjacent channel. After 24 hours of co-culture, cells were primarily retained within their respective channels.

[0084] In preliminary 2D experiments, it was found that L929 fibroblasts alone did not activate RAW264.7 macrophages, and the effects of lipopolysaccharide (LPS) stimulation on macrophage morphology and activation were confirmed. These measurements were then projected to a three-dimensional microfluidic system. The effects of cell count and incubation time on B16-F10 melanoma spheroid formation were also determined. The information was used to select the optimal parameters to form spheroids for use in a microfluidic device. F-actin staining of fibroblasts was also performed.

[0085] Device characterization results and data for microfluidic co-culture of fibroblasts and macrophages with melanoma spheroids and alginate-based biomaterials are projected.

[0086] Example 3

[0087] During the experiment, observe different sizes of fluorescently labeled dextran molecules diffuse into the central channel of microfluidic device from lateral (lateral) channel. Experiment with 10k molecular weight (MW) dextran. Also experiment with 10k, 70k MW dextran. Measure dextran diffusion at zero hour, three hours and twenty-four hours. Molecular weight is that the dextran molecule of 10k and 70k successfully diffuses from lateral channel (from second channel or third channel towards first (center) channel) across the channel of matrigel filling of device in 24 hours. Across the whole device, observe the linear gradient (measured by fluorescence intensity) of dextran concentration. These experiments have confirmed the possibility of producing cytokine gradient in device.

[0088] Example 4

[0089] Macrophages and fibroblasts were co-cultured in the lateral channels and the central channel of the microfluidic device, respectively, and a suture was provided in the central channel during the co-culture to monitor any development of fibrosis on the suture.Suture can optionally be placed in the microfluidic device during assembly of the device.

[0090] Example 5

[0091] Melanoma spheroids were developed for placement in microfluidic devices. Melanoma spheroid diameters were measured as a function of cell density and incubation time. Live-dead staining was performed on melanoma spheroids formed from 8 cells / μL (i), 27 cells / μL (ii), and 54 cells / μL (iii) after 72 hours of incubation. Live and dead cells were marked with differentiating colors.

[0092] Example 6

[0093] The placement of biomaterials in microfluidic devices was studied. Alginate beads were implanted in the microfluidic device implantation holes (recesses / containers). Alginate was selected as a model biomaterial because it is known to cause fibrosis. Sutures were placed in the fibrosis chip. Matrigel was successfully gelled around the sutures. These results confirm the feasibility of biomaterials being placed in the fibrosis chip. Even without incorporating implantation holes, it is possible to place sutures in the device.

[0094] Example 7

[0095] Studying Fibroblast-Induced Collagen Deposition in a Two-Dimensional Model. To determine whether L929 fibroblasts deposit collagen in response to stimuli, a well plate experiment was performed according to the protocol described in Good et al., "A High Content. Phenotypic 'Scar-In-A-Jar' Assay for Rapid Quantification of Collagen Fibrillogenesis Using Disease-Derived Pulmonary Fibroblasts," BMC Biomedical Engineering, 1: 14, 1-10 (2019), which is incorporated herein by reference in its entirety. Exposure to a 2-dose containing L-ascorbic acid, Immunostaining experiments were performed on L929 cells and primary mouse skin fibroblasts in culture medium containing TGF-β1 and TGF-β1. It was observed that primary cells responded to TGF-β1 treatment with increased collagen I deposition and α-SMA expression. In contrast, L929 cells did not have α-SMA expression and did not respond to TGF-β1 treatment with increased collagen I deposition. Additionally, the type of L-ascorbic acid used had some effect on collagen I deposition. These findings indicate the correlation and effect of cell type and L-ascorbic acid type on extracellular matrix deposition in vitro.

[0096] Example 8

[0097] One method that can be used to visualize and quantify collagen deposition in microfluidic devices is picrosirius red staining. Picrosirius red staining of paraformaldehyde-fixed decellularized extracellular matrix deposited on 2D surfaces by WI-38 human lung cells shows collagen deposition ( Fig.11 ). Fig.11, Part A shows the results of one day and nine days after plating cells on Permanox plastic four-chamber slides (250,000 cells / chamber). Cells were grown in chambers with or without fibronectin coating and with or without L-ascorbic acid. Cells grown in Permanox plastic 4-chamber slides (250,000 cells / chamber) showed an increase in collagen deposition in the case of L-ascorbic acid treatment and in the case of longer culture time (nine days versus one day). In the absence of fibronectin coating, no visible collagen deposition remained after 14-15 minutes of decellularization. Fig.11 , Part B shows the results of plated WI-38 cells (44,444 cells / well) grown in 96-well plates nine days after plating with a five-minute decellularization time. Even in the absence of a fibronectin coating, the cells appear to deposit retained collagen fibers with a short decellularization time of five minutes. Fig.11 , Part C shows the results of plated WI-38 cells (97,222 cells / chamber) grown in eight chamber glass slides nine days after plating with a five minute decellularization time. Even in the absence of a fibronectin coating, the cells appear to deposit retained collagen fibers with a short decellularization time of five minutes.

[0098] Example 9

[0099] The use of different materials within microfluidic devices is of interest because it has been found that cells sometimes behave differently on plastic surfaces compared to glass surfaces ( Figure 12-17 In addition to any materials placed in the device, the materials used to make the microfluidic device can also play a role in how the cells will behave. Treatment of the cells after culture (e.g., fixation methods and decellularization) can also affect the visualization of proteins of interest such as collagen I and α-smooth muscle actin (α-SMA). Figure 12-17 ). A variety of different treatment and staining methods can be used with microfluidic devices.

[0100] Deposition of collagen I and α-SMA by WI-38 cells in 96-well plates was studied. Cells were plated at 44,444 cells / well and grown in medium including L-ascorbic acid. Cells were grown in wells with no coating or fibronectin coating. Cells were grown with or without 5 ng / mL TGF-β1. Fig.12 ) or methanol ( Fig.13 ) Fixed cells. "Merged" fluorescent images of collagen I, α-SMA and cell nuclei are shown in consecutive columns. Images are shown at 20X magnification. Fig.14 and 15Quantitative fluorescence intensity of collagen I and α-SMA channels are shown. Image quantification was performed in ImageJ, where the average image intensity for each channel was calculated. Statistical analysis was performed in GraphPad Prism using 2-way ANOVA with multiple comparisons. The significant difference between the collagen I intensity in the PFA-fixed wells relative to the methanol-fixed wells was obvious at low magnifications, but not at higher magnifications. In contrast, the significant difference between the α-SMA intensity in the PFA-fixed wells relative to the methanol-fixed wells was obvious at all magnifications tested. Fig.16 and Fig.17 Shows something like Fig.12 and Fig.13 The results of the cell study of the cells shown in (but the cells were grown in eight chamber glass slides). The cells were plated at a concentration of 97,222 cells / chamber for conventional culture medium treatment, and at a concentration of 18,421 cells / chamber for Ficoll culture medium treatment. Again, compared with PFA-fixed cells, an improvement in staining was observed in methanol-fixed cells. Additionally, collagen I seems to be fibrous in methanol-fixed cells, but seems to be located around the nucleus in PFA-fixed cells.

[0101] The present disclosure may include any combination of these various features or embodiments as shown above and / or below as sentences and / or paragraphs.Any combination of features disclosed herein is considered part of the present disclosure.

[0102] 1. A fibrosis diagnostic system, comprising:

[0103] A microfluidic device, comprising:

[0104] a device housing, the device housing containing the device interior;

[0105] A plurality of channels within the interior of the device, the plurality of channels comprising:

[0106] a first channel configured to receive a target object, a matrix, and a first biological composition configured to promote fibrosis of the target object,

[0107] a second channel configured to receive a second biological composition configured to promote fibrosis in the target subject,

[0108] a first diaphragm separating the first channel from the second channel and comprising a first plurality of openings providing fluid communication between the first channel and the second channel,

[0109] a third channel configured to receive a third biological composition configured to promote fibrosis in the target subject, and

[0110] a second diaphragm separating the first channel from the third channel and comprising a second plurality of openings providing fluid communication between the first channel and the third channel; and

[0111] A plurality of channel ports in the device housing are connected to the plurality of channel fluids, the plurality of channel ports comprising a first group of channel ports connected to the first channel fluid, a second group of channel ports connected to the second channel fluid, and a third group of channel ports connected to the third channel fluid, wherein each group of channel ports comprises one or more channel ports.

[0112] 2. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the system is configured to form a gradient of at least one type of molecule across the first channel, the second channel, or the third channel, or any combination thereof.

[0113] 3. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the at least one type of molecule comprises a chemotactic molecule.

[0114] 4. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the chemotactic molecule promotes the movement of at least one cell type present in the first biological composition, the second biological composition, or the third biological composition, or any combination thereof.

[0115] 5. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the first biological composition, the second biological composition, or the third biological composition, or any combination thereof, comprises the at least one type of molecule.

[0116] 6. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the second group of channel ports and the third group of channel ports each comprise two or more channel ports.

[0117] 7. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein each group of channel ports comprises two or more channel ports.

[0118] 8. A fibrosis diagnostic system according to any of the preceding or following embodiments / features / aspects, wherein the second group of channel ports and the third group of channel ports each include a first channel port and a second channel port, the first channel port being close to the first end of its corresponding channel, and the second channel port being close to the second end of its corresponding channel.

[0119] 9. A fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein each group of channel ports in the plurality of groups of channel ports comprises a first channel port and a second channel port, the first channel port being proximate to a first end of its corresponding channel, and the second channel port being proximate to a second end of its corresponding channel.

[0120] 10. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the first channel comprises a central channel port configured to receive the target object into the first channel.

[0121] 11. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the central channel port is between the first end of the first channel and the second end of the second channel.

[0122] 12. A fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein one or more of the plurality of channel ports comprises a seal, a valve or a closure or any combination thereof, wherein the seal, the valve or the closure or any combination thereof is configured to move between an open state and a closed state.

[0123] 13. A fibrosis diagnostic system according to any of the preceding or following embodiments / features / aspects, comprising a retaining element consisting of one or more of the first channel, the second channel, and the third channel, wherein the retaining element is configured to retain at least one component that can be inserted into one or more of the first channel, the second channel, and the third channel.

[0124] 14. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the retaining element comprises a modification of the channel surface.

[0125] 15. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the modification comprises a biomolecule.

[0126] 16. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the biomolecule comprises a polypeptide, a polysaccharide, a lipid or a polynucleotide or any combination thereof.

[0127] 17. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the modification comprises a polymer, a metal or a ceramic or any combination thereof.

[0128] 18. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the device housing comprises a polymer, a metal or a ceramic or any combination thereof.

[0129] 19. The fibrosis diagnostic system of any preceding or following embodiment / feature / aspect, wherein the device housing comprises a first polymer and the modification comprises a second polymer, the first polymer and the second polymer being different from each other in chemical composition.

[0130] 20. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the modification comprises a physical structure.

[0131] 21. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the physical structure and the device housing comprise a common material.

[0132] 22. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the modification comprises a container for receiving the target object.

[0133] 23. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the container comprises a recess in the channel surface.

[0134] 24. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the physical structure comprises a plurality of physical structures.

[0135] 25. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the plurality of physical structures comprises a plurality of recesses in the channel surface or a plurality of protrusions from the channel surface, or both.

[0136] 26. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the first channel, the second channel or the third channel or any combination thereof comprises a curved cross section or a straight cross section or both.

[0137] 27. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the first channel, the second channel or the third channel or any combination thereof comprises a constant width or a constant diameter or both.

[0138] 28. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the first channel, the second channel or the third channel or any combination thereof comprises a variable width or a constant diameter or both.

[0139] 29. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein two or more channels of the plurality of channels have a common length.

[0140] 30. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein two or more channels of the plurality of channels are parallel to each other.

[0141] 31. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the first channel is between the second channel and the third channel.

[0142] 32. A fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein two or more of the first channel, the second channel and the third channel differ from each other in volume, surface area, cross-sectional area, length, width or diameter, or any combination thereof.

[0143] 33. A fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the width, diameter, or both of the first channel are greater than the width, diameter, or both of the second channel or the third channel or both.

[0144] 34. A fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the cross-sectional area, width, diameter or any combination thereof of one or more of the first channel, the second channel and the third channel differ along their length.

[0145] 35. A fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein one or more of the first channel, the second channel and the third channel comprises a first peripheral segment near the first end of the channel, a second peripheral segment near the second end of the channel, and a central segment between the first peripheral segment and the second peripheral segment.

[0146] 36. A fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the central segment comprises a cross-sectional area, width or diameter, or any combination thereof, that is greater than the cross-sectional area, width or diameter, or any combination thereof, of one or both of the first peripheral segment and the second peripheral segment.

[0147] 37. A fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the central segment of the first channel comprises a cross-sectional area, width or diameter, or any combination thereof, that is larger than the cross-sectional area, width or diameter, or any combination thereof, of one or both of the first peripheral segment and the second peripheral segment of the first channel.

[0148] 38. A fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein one or both of the first peripheral segment and the second peripheral segment are collinear with the central segment relative to their respective central longitudinal axes.

[0149] 39. A fibrosis diagnostic system according to any of the preceding or following embodiments / features / aspects, wherein one or both of the first peripheral segment and the second peripheral segment form an angle other than 180° with respect to their respective central longitudinal axes with the central segment.

[0150] 40. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the angle comprises a right angle, an acute angle, or an obtuse angle, or any combination thereof.

[0151] 41. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein two or more of the plurality of channels are located in a common plane.

[0152] 42. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the first channel, the second channel and the third channel of the plurality of channels are located in a common plane.

[0153] 43. A fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the length of the first diaphragm is less than the length of the first channel or the length of the second channel or both.

[0154] 44. A fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the length of the second diaphragm is less than the length of the first channel or the length of the third channel or both.

[0155] 45. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the first diaphragm, the second diaphragm, or both comprise a membrane.

[0156] 46. ​​The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the membrane comprises a porous membrane.

[0157] 47. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the membrane comprises a semipermeable membrane.

[0158] 48. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the membrane comprises a biological membrane or a synthetic membrane or both.

[0159] 49. A fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the first diaphragm comprises a first plurality of columns separated by the first plurality of openings, the second diaphragm comprises a second plurality of columns separated by the second plurality of second openings, or both.

[0160] 50. The fibrosis diagnostic system of any preceding or following embodiment / feature / aspect, wherein the first plurality of columns or the second plurality of columns or both have a straight cross-section, a curved cross-section or both.

[0161] 51. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the first plurality of columns or the second plurality of columns or both are trapezoidal.

[0162] 52. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein:

[0163] The openings in the first plurality of openings differ in number, size, shape or spacing, or any combination thereof; or

[0164] The openings in the second plurality of openings differ in number, size, shape or spacing, or any combination thereof; or

[0165] Both.

[0166] 53. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein:

[0167] The openings in the first plurality of openings vary in number, size, shape or spacing, or any combination thereof, along one or more dimensions of the first diaphragm; or

[0168] The openings in the second plurality of openings vary in number, size, shape or spacing, or any combination thereof, along one or more dimensions of the second diaphragm; or

[0169] Both.

[0170] 54. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the first plurality of openings and the second plurality of openings are different from each other.

[0171] 55. The fibrosis diagnostic system of any preceding or following embodiment / feature / aspect, wherein the device housing comprises a substrate, and the plurality of channels and the plurality of channel ports are formed in the substrate.

[0172] 56. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the device housing comprises silicone.

[0173] 57. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the device housing comprises polydimethylsiloxane (PDMS).

[0174] 58. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the device housing comprises a flexible material or a rigid material or both.

[0175] 59. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the device housing comprises a conductor or an insulator or both.

[0176] 60. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the substrate comprises a first material and one or more channel surfaces comprise a second material different from the first material.

[0177] 61. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the second material is different from the contents of one or more channels in which the second material is present.

[0178] 62. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the second material is a coating on the first material.

[0179] 63. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein one or more channel surfaces are coated with a plurality of different materials.

[0180] 64. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein one or more channel surfaces are coated with a plurality of coatings.

[0181] 65. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein at least two of the plurality of coatings differ in material composition.

[0182] 66. A fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the substrate comprises a first material, and the multiple coatings comprise a first coating adjacent to the substrate on one or more channel surfaces and a second coating covering the first coating, the first coating comprising a second material, and the second coating comprising a third material.

[0183] 67. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the first material and the second material are different from each other, and the second material and the third material are different from each other.

[0184] 68. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the first material, the second material and the third material are different from each other.

[0185] 69. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the plurality of coatings comprises at least two layers having average thicknesses different from each other.

[0186] 70. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the material composition, thickness or patterning or a combination thereof of the coating on the surface of one or more channels varies along the length of the one or more channels.

[0187] 71. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein two or more of the channels differ in the material composition of their surface coatings.

[0188] 72. A fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the first channel comprises a first coating, the second channel comprises a second coating, and the third channel comprises a third coating; the first coating, the second coating and the third coating differ in material composition, thickness or patterning or a combination thereof.

[0189] 73. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the first channel, the second channel and the third channel are coated with the same material.

[0190] 74. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein one or more coatings comprise organic material or inorganic material or both.

[0191] 75. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein one or more coatings comprise a polymer.

[0192] 76. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein one or more coatings comprise biomolecules.

[0193] 77. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein one or more coatings comprise proteins or nucleic acids, lipids or carbohydrates or any combination thereof.

[0194] 78. A fibrosis diagnostic system according to any of the preceding or following embodiments / features / aspects, wherein one or more coatings comprise fibronectin, an extracellular matrix-derived composition, a basement membrane extract, a decellularized extracellular matrix, a dissolved basement membrane preparation, a mixture of extracellular matrix proteins, a tumor-derived composition, matrigel or hyaluronic acid or any combination thereof.

[0195] 79. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the substrate comprises silicon and one or more channel surfaces are coated with a coating comprising silane.

[0196] 80. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein one or more channel surfaces are subjected to silanization.

[0197] 81. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein one or more channel surfaces are treated with plasma.

[0198] 82. The fibrosis diagnostic system of any preceding or following embodiment / feature / aspect, wherein the plasma comprises oxygen.

[0199] 83. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein one or more coatings are applied by: emerging, spraying, brushing, injecting, precipitating, polymerizing or vapor deposition or any combination thereof.

[0200] 84. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein one or more channel surfaces comprise a laminate material.

[0201] 85. A fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the device housing comprises a transparent material, a translucent material, a colored material, a polarized material, a material that selectively transmits a wavelength of electromagnetic radiation, a material that selectively refracts a wavelength of electromagnetic radiation, a material that selectively reflects a wavelength of electromagnetic radiation, or an opaque material, or any combination thereof.

[0202] 86. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the device housing comprises a window allowing a view of the interior of the device.

[0203] 87. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the view is a view of one or more channels of the plurality of channels.

[0204] 88. The fibrosis diagnostic system of any preceding or following embodiment / feature / aspect, wherein the view comprises a view of the first channel or the target object or both.

[0205] 89. The fibrosis diagnostic system of any preceding or following embodiment / feature / aspect, wherein the device housing comprises a plurality of windows allowing for a plurality of views of the interior of the device.

[0206] 90. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the target object comprises organic material or inorganic material or both.

[0207] 91. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the target object comprises a tissue or a medical device or both.

[0208] 92. A fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the target object comprises a spheroid, an organoid, an alginate-based biomaterial, a scaffold-supported cell structure or a bioprinted structure or any combination thereof.

[0209] 93. A fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the target object comprises tumor cells, malignant cells or benign cells or any combination thereof.

[0210] 94. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the target comprises one or more coatings.

[0211] 95. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the one or more coatings of the target are different from the matrix.

[0212] 96. A fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the one or more coatings of the target are different from the one or more coatings of the one or more channel surfaces in terms of material composition, thickness or patterning, or a combination thereof.

[0213] 97. A fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the one or more coatings of the target are the same as the one or more coatings of the one or more channel surfaces in terms of material composition, thickness or patterning or a combination thereof.

[0214] 98. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the target is coated with a plurality of different materials.

[0215] 99. The fibrosis diagnostic system of any preceding or following embodiment / feature / aspect, wherein the target is coated with multiple coatings.

[0216] 100. The fibrosis diagnostic system of any preceding or following embodiment / feature / aspect, wherein at least two of the plurality of coatings differ in material composition, thickness or patterning, or a combination thereof.

[0217] 101. A fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the multiple coatings comprise a first coating adjacent to the target and a second coating covering the first coating, the first coating comprising a first material, and the second coating comprising a second material.

[0218] 102. The fibrosis diagnostic system of any preceding or following embodiment / feature / aspect, wherein the target is coated prior to being placed in the microfluidic device.

[0219] 103. The fibrosis diagnostic system of any preceding or following embodiment / feature / aspect, wherein the matrix comprises a hydrogel.

[0220] 104. A fibrosis diagnostic system according to any of the preceding or following embodiments / features / aspects, wherein the matrix comprises fibronectin, an extracellular matrix-derived composition, a basement membrane extract, a decellularized extracellular matrix, a dissolved basement membrane preparation, a mixture of extracellular matrix proteins, a tumor-derived composition, matrigel or hyaluronic acid or any combination thereof.

[0221] 105. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the matrix comprises a natural polymer, an artificial polymer or a cross-linked polymer or any combination thereof.

[0222] 106. A fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the matrix comprises a polysaccharide, a polypeptide, a lipid, a polynucleotide, hyaluronic acid, hyaluronic acid methacrylate, hyaluronic acid diacrylate, fibronectin, fibrinogen, collagen or methacrylate collagen or any combination thereof.

[0223] 107. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the first biological composition comprises fibroblasts or endothelial cells or both.

[0224] 108. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the first biological composition comprises serum, ascorbic acid, retinoic acid, proline or bleomycin or any combination thereof.

[0225] 109. A fibrosis diagnostic system according to any of the preceding or following embodiments / features / aspects, wherein the first biological composition comprises biomolecules that promote fibrosis, complement proteins, coagulation-related proteins, bacterial cell wall proteins or lipopolysaccharide (LPS) or any combination thereof.

[0226] 110. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the second biological composition comprises at least one type of biological cell capable of promoting fibrosis.

[0227] 111. A fibrosis diagnostic system according to any of the preceding or following embodiments / features / aspects, wherein the second biological composition comprises factors that promote the differentiation of biological cells into biological cell types capable of promoting fibrosis, factors that promote the expression of biological molecules that promote fibrosis, or factors that promote the expansion of biological cell types capable of promoting fibrosis, or any combination thereof.

[0228] 112. A fibrosis diagnostic system according to any of the preceding or following embodiments / features / aspects, wherein the second biological composition comprises serum, platelets, leukocytes, macrophages, monocytes, peripheral blood mononuclear cells (PBMC), stem cells, mast cells, dendritic cells, neutrophils, granulocytes, T cells or B cells or any combination thereof.

[0229] 113. A fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the third biological composition comprises a population of molecules capable of forming a gradient radiating from the third channel into the first channel or into both the first channel and the second channel.

[0230] 114. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the third biological composition comprises a cytokine, a chemokine, a cell nutrient, a transcription factor or a growth factor or any combination thereof.

[0231] 115. A fibrosis diagnostic system according to any of the preceding or following embodiments / features / aspects, wherein the third biological composition comprises CCL-2 / MCP-1, CCL3, CCL11, fibroblast growth factor (FGF), interferon, IFN-γ, interleukin (IL), IL-4, IL-1α, IL-1β, IL-2, IL-5, IL-6, IL-7, IL-9, IL-13, IL-17A, IL-17F, IL-21, IL-22, IL-25, platelet-derived growth factor (PDGF), TGF-α, TGF-β, granulocyte colony stimulating factor (G-CSF), macrophage colony stimulating factor (M-CSF), granulocyte macrophage colony stimulating factor (GM-CSF), prothrombin or thrombin or any combination thereof.

[0232] 116. A fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the at least one type of molecule of the gradient comprises a cytokine, a chemokine, a cell nutrient, a transcription factor or a growth factor or any combination thereof.

[0233] 117. The fibrosis diagnostic system of any preceding or following embodiment / feature / aspect, wherein said at least one type of molecule of said gradient comprises transforming growth factor β (TGF-β).

[0234] 118. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the TGF-β comprises TGF-β1.

[0235] 119. A fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the first biological composition, the second biological composition, or the third biological composition, or any combination thereof, comprises labeled cells or labeled biomolecules or both.

[0236] 120. A fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the first biological composition, the second biological composition or the third biological composition or any combination thereof comprises a compound with known or potential anti-fibrotic efficacy.

[0237] 121. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, further comprising the matrix, the first biological composition, the second biological composition or the third biological composition or any combination thereof.

[0238] 122. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, further comprising the target object.

[0239] 123. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, further comprising a gradient of at least one molecule across the first channel, the second channel, or the third channel, or any combination thereof.

[0240] 124. A fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein one or more components of the matrix, the first biological composition, the second biological composition or the third biological composition or any combination thereof are anchored to a channel surface.

[0241] 125. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, further comprising a detector comprising a sensor configured to detect fibrosis of the target object or a characteristic indicating fibrosis of the target object.

[0242] 126. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the detector is configured to receive the microfluidic device.

[0243] 127. The fibrosis diagnostic system of any preceding or following embodiment / feature / aspect, wherein the detector comprises a detector housing including a detector interior.

[0244] 128. A fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the detector further comprises a receiving portion configured to receive the microfluidic device, the receiving portion comprising an opening in the detector housing.

[0245] 129. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, further comprising a thermal subsystem.

[0246] 130. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the thermal subsystem comprises a heater or a cooler or both.

[0247] 131. A fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the thermal subsystem is configured to regulate the temperature of the microfluidic device.

[0248] 132. The fibrosis diagnostic system of any preceding or following embodiment / feature / aspect, wherein the detector comprises the thermal subsystem.

[0249] 133. A fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the thermal subsystem is configured to regulate the temperature of the microfluidic device when the microfluidic device is in the receiving portion.

[0250] 134. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, further comprising a fluid subsystem configured to be in fluid communication with the microfluidic device.

[0251] 135. A fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the fluid subsystem is configured to load one or more components into the microfluidic device.

[0252] 136. A fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the fluid subsystem is configured to supply solid, liquid or gas or any combination thereof to the microfluidic device.

[0253] 137. A fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the fluid subsystem is configured to load the target object, the matrix, the first biological composition, the second biological composition or the third biological composition or any combination thereof into the microfluidic device.

[0254] 138. The fibrosis diagnostic system of any preceding or following embodiment / feature / aspect, wherein the fluid subsystem is configured to supply one or more nutrients to the microfluidic device.

[0255] 139. The fibrosis diagnostic system of any preceding or following embodiment / feature / aspect, wherein the fluid subsystem is configured to regulate the pressure within the interior of the microfluidic device.

[0256] 140. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the fluid subsystem is configured to adjust a gradient across the first channel, the second channel, or the third channel, or any combination thereof.

[0257] 141. The fibrosis diagnostic system of any preceding or following embodiment / feature / aspect, wherein the fluid subsystem is configured to remove one or more components from the microfluidic device.

[0258] 142. A fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the detector comprises the fluid subsystem.

[0259] 143. A fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the fluid subsystem is configured to be in fluid communication with the microfluidic device when the microfluidic device is in the receiving portion.

[0260] 144. A fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the sensor is configured to detect particles or waves or both indicative of fibrosis emitted from the target object or the microfluidic device or both.

[0261] 145. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the sensor is configured to detect electromagnetic radiation.

[0262] 146. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the electromagnetic radiation comprises ultraviolet radiation, visible light or infrared radiation or any combination thereof.

[0263] 147. A fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the electromagnetic radiation comprises fluorescence or phosphorescence or both.

[0264] 148. A fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the detector further comprises a particle source or a wave source or both, wherein the particle source or the wave source or both are configured to transmit particles, waves or both to the target object or the microfluidic device or both.

[0265] 149. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, further comprising an electromagnetic radiation source.

[0266] 150. The fibrosis diagnostic system of any preceding or following embodiment / feature / aspect, wherein the detector comprises the electromagnetic radiation source.

[0267] 151. A fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the electromagnetic radiation source comprises a light emitting diode, a laser, an incandescent light source, an arc light source, a halogen light source, a chemical vapor light source or a fluorescent light source or any combination thereof.

[0268] 152. A fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the detector further comprises an optical subsystem configured to transmit radiation or affect radiation or both.

[0269] 153. A fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the optical subsystem comprises one or more lenses, one or more filters, one or more mirrors or one or more diffraction gratings or any combination thereof.

[0270] 154. A fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the optical subsystem or the sensor or both comprise a microscope or one or more components thereof.

[0271] 155. A fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the optical subsystem comprises the sensor or the electromagnetic radiation source or both.

[0272] 156. A fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the sensor is configured to generate an output signal based on an input signal detected by the sensor.

[0273] 157. A fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the detector further comprises a transmitter, a receiver, a transceiver or any combination thereof, and the transmitter, the receiver, the transceiver or any combination thereof are configured to receive information or send information or both.

[0274] 158. A fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the information comprises the output signal.

[0275] 159. A fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the information comprises an operating signal for operating the sensor.

[0276] 160. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the detector further comprises a data port.

[0277] 161. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, further comprising a processor configured to receive and process information.

[0278] 162. A fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the processor is configured to control one or more subsystems or components of the fibrosis diagnostic system.

[0279] 163. A fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the processor is configured to control the sensor, the electromagnetic radiation source, the optical subsystem, the thermal subsystem or the fluid subsystem or any combination thereof.

[0280] 164. A fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the processor is configured to receive the output signal from the sensor, receive an operation signal from a user input, or both.

[0281] 165. A fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the detector comprises the processor.

[0282] 166. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, further comprising a memory configured to communicate with the processor.

[0283] 167. A fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the memory is configured to store a program for controlling the fibrosis diagnostic system or components thereof.

[0284] 168. A fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the processor comprises the memory.

[0285] 169. A fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the detector comprises the memory.

[0286] 170. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, further comprising a user interface.

[0287] 171. A fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the user interface comprises a monitor, a button, a keyboard, a mouse, a light signal, a camera, an eyepiece lens, a speaker or a microphone or any combination thereof.

[0288] 172. A fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the detector comprises the user interface.

[0289] 173. A fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, further comprising a power supply or a power supply interface or both.

[0290] 174. A fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, wherein the detector comprises the power supply or the power supply interface or both.

[0291] 175. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, further comprising a kit comprising the microfluidic device and one or more reagents for use in the microfluidic device.

[0292] 176. The fibrosis diagnostic system according to any preceding or following embodiment / feature / aspect, further comprising a kit comprising:

[0293] The microfluidic device; and

[0294] The matrix, the first biological composition, the second biological composition, or the third biological composition, or any combination thereof.

[0295] 177. A method of analyzing a target subject's susceptibility to fibrosis, the method comprising:

[0296] placing a target object, a matrix, and a first biological composition in a first channel of the microfluidic device according to any preceding or following embodiment / feature / aspect;

[0297] placing a second biological composition in a second channel of the microfluidic device;

[0298] placing a third biological composition in a third channel of the microfluidic device; and

[0299] The target object or the first channel or both are analyzed to determine the presence or absence of fibrosis associated with the target object.

[0300] 178. A method according to any preceding or following embodiment / feature / aspect, wherein the analysis is performed after a predetermined time period after placing the target object, the matrix, the first biological composition, the second biological composition, or the third biological composition, or any combination thereof, in the microfluidic device.

[0301] 179. A method according to any preceding or following embodiment / feature / aspect, wherein the predetermined time period is sufficient to allow a gradient of at least one type of molecule to form.

[0302] 180. The method of any preceding or following embodiment / feature / aspect, wherein the analysis is performed two or more times.

[0303] 181. A method according to any preceding or following embodiment / feature / aspect, wherein any two consecutive analyses are separated by a predetermined time period.

[0304] 182. A method according to any preceding or following embodiment / feature / aspect, wherein the analysis comprises removing one or more components from the microfluidic device.

[0305] 183. A method according to any preceding or following embodiment / feature / aspect, wherein the analysis comprises an immunoassay.

[0306] 184. A method according to any preceding or following embodiment / feature / aspect, wherein the analysis comprises the use of antibodies.

[0307] 185. A method according to any preceding or following embodiment / feature / aspect, wherein the antibody is labeled.

[0308] 186. A method according to any preceding or following embodiment / feature / aspect, wherein the analysis comprises applying a primary antibody and a secondary antibody comprising a detectable label and binding to the primary antibody.

[0309] 187. A method according to any preceding or following embodiment / feature / aspect, wherein the primary antibody binds to collagen, α-smooth muscle actin, or both.

[0310] 188. A method according to any preceding or following embodiment / feature / aspect, wherein the analysis comprises a histological assay.

[0311] 189. The method of any preceding or following embodiment / feature / aspect, wherein the analyzing comprises applying a picrosirius red dye.

[0312] 190. The method of any preceding or following embodiment / feature / aspect, wherein the analyzing comprises visually inspecting the target object or the first channel or both for one or more indications of fibrosis associated with the target object.

[0313] 191. A method according to any preceding or following embodiment / feature / aspect, wherein the analyzing comprises the use of microscopy.

[0314] 192. A method according to any preceding or following embodiment / feature / aspect, wherein the analysis comprises use of one or more spectroscopic techniques.

[0315] 193. A method according to any preceding or following embodiment / feature / aspect, wherein the analysis comprises nonlinear optical (NLO) multiphoton microscopy, second harmonic generation (SHG) imaging, third harmonic generation (THG) imaging, two-photon excitation fluorescence (TPEF), two-photon excitation fluorescence (TPEF), cell tracker dye imaging, visible light imaging, (ELISA), Gliese assay, histopathology, immunostaining, cleared tissue microscopy, or proteomic profiling of homogenized hydrogels, or any combination thereof.

[0316] 194. A method according to any preceding or following embodiment / feature / aspect, wherein the analysis comprises using a detector to detect one or more indicators of fibrosis associated with the target object.

[0317] 195. A method according to any preceding or following embodiment / feature / aspect, wherein the analysis comprises inserting the microfluidic device into a receiving portion.

[0318] 196. The method according to any preceding or following embodiment / feature / aspect, further comprising preparing the target object, the matrix, the first biological composition, the second biological composition or the third biological composition or any combination thereof from a patient sample.

[0319] 197. The method according to any preceding or following embodiment / feature / aspect, further comprising implanting the target object into or on a patient.

[0320] 198. The method of any preceding or following embodiment / feature / aspect, further comprising implanting a medical device or tissue or both into or on a patient, wherein the target object is a representative model of the medical device or tissue or both.

[0321] 199. The method of any preceding or following embodiment / feature / aspect, further comprising removing a medical device or tissue or both from a patient, wherein the target object is a representative model of the medical device or tissue or both.

[0322] 200. The method according to any preceding or following embodiment / feature / aspect, further comprising administering an anti-fibrotic agent to the patient.

[0323] 201. A method according to any preceding or following embodiment / feature / aspect, wherein the analysis comprises determining a quantitative measure of fibrosis.

[0324] 202. The method of any preceding or following embodiment / feature / aspect, wherein the dose of anti-fibrotic agent administered to the patient is based on a quantitative measure of the fibrosis.

[0325] 203. A method according to any preceding or following embodiment / feature / aspect, wherein the anti-fibrotic agent is administered to the patient before, simultaneously with, or after implantation of the target object, a tissue associated with the target object, or a medical device associated with the target object, or any combination thereof.

[0326] 204. The method according to any preceding or following embodiment / feature / aspect, further comprising administering to the patient one or more therapeutic agents in addition to or instead of the anti-fibrotic agent.

[0327] 205. The method of any preceding or following embodiment / feature / aspect, wherein the patient is a mammalian patient or a human patient or both.

[0328] 206. A method for screening a therapy for efficacy in treating fibrosis, the method comprising:

[0329] placing a target object, a matrix, and a first biological composition in a first channel of the microfluidic device according to any preceding or following embodiment / feature / aspect;

[0330] placing a second biological composition in a second channel of the microfluidic device;

[0331] placing a third biological composition in a third channel of the microfluidic device;

[0332] performing the therapy on the microfluidic device;

[0333] analyzing the target object or the first channel or both to determine the presence, absence or extent of fibrosis associated with the target object; and

[0334] The efficacy of the therapy is determined based on the presence, absence, or difference in the extent of fibrosis in the absence of the therapy.

[0335] 207. A method according to any preceding or following embodiment / feature / aspect, wherein the therapy comprises a therapeutic compound, and the performing comprises placing the therapeutic compound in the first channel, the second channel, or the third channel, or any combination thereof.

[0336] 208. A method according to any preceding or following embodiment / feature / aspect, wherein the therapeutic compound comprises a drug compound or a biologic or both.

[0337] 209. The method of any preceding or following embodiment / feature / aspect, wherein the therapy comprises modification of the target subject.

[0338] 210. A method according to any preceding or following embodiment / feature / aspect, wherein the therapy comprises placing a medical device in the first channel, the second channel, or the third channel, or any combination thereof.

[0339] 211. A method according to any preceding or following embodiment / feature / aspect, wherein the therapy comprises applying radiation to the microfluidic device.

[0340] 212. The method of any preceding or following embodiment / feature / aspect, wherein the therapy further comprises treating the patient with a selected therapy.

[0341] 213. A method for screening a therapy for efficacy in treating cancer, the method comprising:

[0342] placing a target object, a matrix, and a first biological composition in a first channel of the microfluidic device according to any preceding or following embodiment / feature / aspect, wherein the target object comprises a tumor cell;

[0343] placing a second biological composition in a second channel of the microfluidic device;

[0344] placing a third biological composition in a third channel of the microfluidic device;

[0345] performing the therapy on the microfluidic device;

[0346] analyzing the target object or the first channel or both to determine the presence, absence or extent of fibrosis associated with the target object; and

[0347] The efficacy of the therapy is determined based on the difference in the presence, absence, or extent of the cancer in the absence of the therapy.

[0348] 214. A non-transitory computer-readable medium storing a program, the program comprising instructions for performing a method according to any preceding or following embodiment / feature / aspect.

[0349] 215. A non-transitory computer-readable medium storing a program, wherein the program comprises instructions for controlling a fibrosis diagnostic system or a subsystem or a component thereof or any combination thereof according to any preceding or following embodiment / feature / aspect.

[0350] The applicant specifically incorporates the full content of all cited references in the present disclosure. Further, when the amount, concentration or other value or parameter is given as a list of range, preferred range or preferred upper limit value and preferred lower limit value, this should be understood as specifically disclosing all ranges formed by any pair of any upper range limit or preferred value and any lower range limit or preferred value, no matter whether the scope is disclosed separately. In the case of narrating numerical range in this article, unless otherwise stated, the scope is intended to include its endpoints, and all integers and fractions in the scope. The scope of the present disclosure is not limited to the specific value narrated when defining the range.

[0351] All documents cited in this specification are incorporated herein by reference as if each document was specifically and individually indicated to be incorporated by reference. The citation of any document is for its disclosure prior to the filing date and should not be construed as an admission that the present disclosure does not entitle the present disclosure to claim any specific subject matter disclosed herein.

[0352] It should be understood that each of the above elements or two or more elements together may also find useful applications in other types of methods different from the above types. Without further analysis, the foregoing will fully reveal the gist of the present disclosure so that others can easily adapt it to various uses while still falling within the scope of the claimed subject matter. The foregoing embodiments are presented only by way of example.

Claims

1. A fibrosis diagnostic system, comprising: A microfluidic device, comprising: a device housing, the device housing containing the device interior; A plurality of channels within the interior of the device, the plurality of channels comprising: a first channel configured to receive a target object, a matrix, and a first biological composition configured to promote fibrosis of the target object, a second channel configured to receive a second biological composition configured to promote fibrosis in the target subject, a first diaphragm separating the first channel from the second channel and comprising a first plurality of openings providing fluid communication between the first channel and the second channel, a third channel configured to receive a third biological composition configured to promote fibrosis in the target subject, and a second diaphragm separating the first channel from the third channel and comprising a second plurality of openings providing fluid communication between the first channel and the third channel; and A plurality of channel ports in the device housing are connected to the plurality of channel fluids, the plurality of channel ports comprising a first group of channel ports connected to the first channel fluid, a second group of channel ports connected to the second channel fluid, and a third group of channel ports connected to the third channel fluid, wherein each group of channel ports comprises one or more channel ports.

2. A fibrosis diagnostic system according to any preceding claim, wherein the system is configured to form a gradient of at least one type of molecule across the first channel, the second channel or the third channel or any combination thereof.

3. A fibrosis diagnostic system according to any preceding claim, wherein the at least one type of molecule comprises a chemotactic molecule.

4. The fibrosis diagnostic system according to any preceding claim, wherein the chemotactic molecule promotes movement of at least one cell type present in the first biological composition, the second biological composition, or the third biological composition, or any combination thereof.

5. The fibrosis diagnostic system according to any preceding claim, wherein the first biological composition, the second biological composition or the third biological composition or any combination thereof comprises the at least one type of molecule.

6. The fibrosis diagnostic system according to any preceding claim, wherein the second group of channel ports and the third group of channel ports each comprise two or more channel ports.

7. A fibrosis diagnostic system according to any preceding claim, wherein each group of channel ports comprises two or more channel ports.

8. A fibrosis diagnostic system according to any preceding claim, wherein the second group of channel ports and the third group of channel ports each include a first channel port and a second channel port, wherein the first channel port is proximate to a first end of its corresponding channel and the second channel port is proximate to a second end of its corresponding channel.

9. A fibrosis diagnostic system according to any preceding claim, wherein each group of channel ports in the plurality of groups of channel ports comprises a first channel port and a second channel port, the first channel port being proximate to a first end of its corresponding channel and the second channel port being proximate to a second end of its corresponding channel.

10. The fibrosis diagnostic system of any preceding claim, wherein the first channel comprises a central channel port configured to receive the target object into the first channel.

11. A fibrosis diagnostic system according to any preceding claim, wherein the central channel port is between a first end of the first channel and a second end of the second channel.

12. A fibrosis diagnostic system according to any preceding claim, wherein one or more of the plurality of channel ports comprises a seal, a valve or a closure or any combination thereof, the seal, the valve or the closure or any combination thereof being configured to move between an open state and a closed state.

13. A fibrosis diagnostic system according to any preceding claim, comprising a retaining element formed by one or more of the first channel, the second channel and the third channel, the retaining element being configured to retain at least one component that can be inserted into one or more of the first channel, the second channel and the third channel.

14. A fibrosis diagnostic system according to any preceding claim, wherein the retaining element comprises a modification of the channel surface.

15. A fibrosis diagnostic system according to any preceding claim, wherein the modification comprises a biomolecule.

16. The fibrosis diagnostic system according to any preceding claim, wherein the biomolecule comprises a polypeptide, a polysaccharide, a lipid or a polynucleotide or any combination thereof.

17. A fibrosis diagnostic system according to any preceding claim, wherein the modification comprises a polymer, a metal or a ceramic or any combination thereof.

18. A fibrosis diagnostic system according to any preceding claim, wherein the device housing comprises a polymer, metal or ceramic or any combination thereof.

19. A fibrosis diagnostic system according to any preceding claim, wherein the device housing comprises a first polymer and the modification comprises a second polymer, the first polymer and the second polymer differing from each other in chemical composition.

20. A fibrosis diagnostic system according to any preceding claim, wherein the modification comprises a physical structure.

21. A fibrosis diagnostic system according to any preceding claim, wherein the physical structure and the device housing comprise a common material.

22. A fibrosis diagnostic system according to any preceding claim, wherein the modification comprises a container for receiving the target object.

23. A fibrosis diagnostic system according to any preceding claim, wherein the container comprises a recess in the channel surface.

24. A fibrosis diagnostic system according to any preceding claim, wherein the physical structure comprises a plurality of physical structures.

25. The fibrosis diagnostic system of any preceding claim, wherein the plurality of physical structures comprises a plurality of recesses in the channel surface or a plurality of protrusions from the channel surface, or both.

26. The fibrosis diagnostic system of any preceding claim, wherein the first channel, the second channel, or the third channel, or any combination thereof, comprises a curvilinear cross section or a linear cross section, or both.

27. The fibrosis diagnostic system of any preceding claim, wherein the first channel, the second channel, or the third channel, or any combination thereof, comprises a constant width or a constant diameter, or both.

28. The fibrosis diagnostic system of any preceding claim, wherein the first channel, the second channel, or the third channel, or any combination thereof, comprises a variable width or a constant diameter, or both.

29. A fibrosis diagnostic system according to any preceding claim, wherein two or more channels of the plurality of channels have a common length.

30. A fibrosis diagnostic system according to any preceding claim, wherein two or more channels of the plurality of channels are parallel to each other.

31. A fibrosis diagnostic system according to any preceding claim, wherein the first channel is between the second channel and the third channel.

32. A fibrosis diagnostic system according to any preceding claim, wherein two or more of the first channel, the second channel and the third channel differ from each other in volume, surface area, cross-sectional area, length, width or diameter, or any combination thereof.

33. The fibrosis diagnostic system of any preceding claim, wherein the first channel has a width, a diameter, or both, greater than the width, a diameter, or both of the second channel or the third channel or both.

34. The fibrosis diagnostic system of any preceding claim, wherein one or more of the first, second and third channels has a cross-sectional area, width, diameter or any combination thereof that varies along its length.

35. A fibrosis diagnostic system according to any preceding claim, wherein one or more of the first channel, the second channel and the third channel comprises a first peripheral segment near the first end of the channel, a second peripheral segment near the second end of the channel, and a central segment between the first peripheral segment and the second peripheral segment.

36. A fibrosis diagnostic system according to any preceding claim, wherein the central segment comprises a cross-sectional area, width or diameter, or any combination thereof, that is greater than the cross-sectional area, width or diameter, or any combination thereof, of one or both of the first and second peripheral segments.

37. A fibrosis diagnostic system according to any preceding claim, wherein the central segment of the first channel comprises a cross-sectional area, width or diameter, or any combination thereof, that is greater than the cross-sectional area, width or diameter, or any combination thereof, of one or both of the first peripheral segment and the second peripheral segment of the first channel.

38. A fibrosis diagnostic system according to any preceding claim, wherein one or both of the first and second peripheral segments are co-linear with the central segment with respect to their respective central longitudinal axes.

39. A fibrosis diagnostic system according to any preceding claim, wherein one or both of the first and second peripheral segments form an angle other than 180° with the central segment relative to their respective central longitudinal axes.

40. The fibrosis diagnostic system according to any preceding claim, wherein the angle comprises a right angle, an acute angle or an obtuse angle or any combination thereof.

41. A fibrosis diagnostic system according to any preceding claim, wherein two or more channels of the plurality of channels lie in a common plane.

42. The fibrosis diagnostic system of any preceding claim, wherein the first channel, the second channel and the third channel of the plurality of channels lie in a common plane.

43. The fibrosis diagnostic system of any preceding claim, wherein the length of the first diaphragm is less than the length of the first channel or the length of the second channel or both.

44. A fibrosis diagnostic system according to any preceding claim, wherein the length of the second diaphragm is less than the length of the first channel or the length of the third channel or both.

45. The fibrosis diagnostic system of any preceding claim, wherein the first diaphragm, the second diaphragm, or both comprise a membrane.

46. ​​A fibrosis diagnostic system according to any preceding claim, wherein the membrane comprises a porous membrane.

47. A fibrosis diagnostic system according to any preceding claim, wherein the membrane comprises a semi-permeable membrane.

48. A fibrosis diagnostic system according to any preceding claim, wherein the membrane comprises a biological membrane or a synthetic membrane or both.

49. The fibrosis diagnostic system of any preceding claim, wherein the first membrane comprises a first plurality of posts separated by the first plurality of openings, the second membrane comprises a second plurality of posts separated by the second plurality of openings, or both.

50. The fibrosis diagnostic system of any preceding claim, wherein the first plurality of pillars or the second plurality of pillars or both have a straight cross-section, a curved cross-section or both.

51. A fibrosis diagnostic system according to any preceding claim, wherein the first plurality of pillars or the second plurality of pillars or both are trapezoidal in shape.

52. A fibrosis diagnostic system according to any preceding claim, wherein: The openings in the first plurality of openings differ in number, size, shape or spacing, or any combination thereof; or The openings in the second plurality of openings differ in number, size, shape or spacing, or any combination thereof; or Both.

53. A fibrosis diagnostic system according to any preceding claim, wherein: The openings in the first plurality of openings vary in number, size, shape or spacing, or any combination thereof, along one or more dimensions of the first diaphragm; or The openings in the second plurality of openings vary in number, size, shape or spacing, or any combination thereof, along one or more dimensions of the second diaphragm; or Both.

54. The fibrosis diagnostic system according to any preceding claim, wherein the first plurality of openings and the second plurality of openings are different from each other.

55. The fibrosis diagnostic system of any preceding claim, wherein the device housing comprises a substrate, and the plurality of channels and the plurality of channel ports are formed in the substrate.

56. A fibrosis diagnostic system according to any preceding claim, wherein the device housing comprises silicone.

57. A fibrosis diagnostic system according to any preceding claim, wherein the device housing comprises polydimethylsiloxane (PDMS).

58. A fibrosis diagnostic system according to any preceding claim, wherein the device housing comprises a flexible material or a rigid material or both.

59. A fibrosis diagnostic system according to any preceding claim, wherein the device housing comprises a conductor or an insulator or both.

60. A fibrosis diagnostic system according to any preceding claim, wherein the substrate comprises a first material and one or more channel surfaces comprise a second material different from the first material.

61. A fibrosis diagnostic system according to any preceding claim, wherein the second material is different from the contents of one or more channels in which the second material is present.

62. A fibrosis diagnostic system according to any preceding claim, wherein the second material is a coating on the first material.

63. A fibrosis diagnostic system according to any preceding claim, wherein one or more channel surfaces are coated with a plurality of different materials.

64. A fibrosis diagnostic system according to any preceding claim, wherein one or more channel surfaces are coated with a plurality of coatings.

65. The fibrosis diagnostic system according to any preceding claim, wherein at least two coatings of the plurality of coatings differ in material composition.

66. A fibrosis diagnostic system according to any preceding claim, wherein the substrate comprises a first material, and the plurality of coatings comprises a first coating adjacent to the substrate on one or more channel surfaces and a second coating covering the first coating, the first coating comprising a second material, the second coating comprising a third material.

67. A fibrosis diagnostic system according to any preceding claim, wherein the first material and the second material are different from each other, and the second material and the third material are different from each other.

68. A fibrosis diagnostic system according to any preceding claim, wherein the first material, the second material and the third material are different from each other.

69. The fibrosis diagnostic system according to any preceding claim, wherein the plurality of coatings comprises at least two layers having average thicknesses different from each other.

70. A fibrosis diagnostic system according to any preceding claim, wherein the material composition, thickness or patterning or a combination thereof of the coating on the surface of one or more channels varies along the length of the one or more channels.

71. A fibrosis diagnostic system according to any preceding claim, wherein two or more of the channels differ in the material composition of their surface coatings.

72. A fibrosis diagnostic system according to any preceding claim, wherein the first channel comprises a first coating, the second channel comprises a second coating, and the third channel comprises a third coating; the first coating, the second coating and the third coating differ in material composition, thickness or patterning or a combination thereof.

73. The fibrosis diagnostic system according to any preceding claim, wherein the first channel, the second channel and the third channel are coated with the same material.

74. A fibrosis diagnostic system according to any preceding claim, wherein one or more coatings comprise organic or inorganic materials or both.

75. A fibrosis diagnostic system according to any preceding claim, wherein one or more coatings comprise a polymer.

76. A fibrosis diagnostic system according to any preceding claim, wherein one or more coatings comprise biomolecules.

77. The fibrosis diagnostic system according to any preceding claim, wherein one or more coatings comprise proteins or nucleic acids, lipids or carbohydrates or any combination thereof.

78. A fibrosis diagnostic system according to any preceding claim, wherein one or more coatings comprise fibronectin, an extracellular matrix-derived composition, a basement membrane extract, a decellularized extracellular matrix, a dissolved basement membrane preparation, a mixture of extracellular matrix proteins, a tumor-derived composition, matrigel or hyaluronic acid, or any combination thereof.

79. A fibrosis diagnostic system according to any preceding claim, wherein the substrate comprises silicon and one or more channel surfaces are coated with a coating comprising silane.

80. A fibrosis diagnostic system according to any preceding claim, wherein one or more channel surfaces are subjected to silanization.

81. A fibrosis diagnostic system according to any preceding claim, wherein one or more channel surfaces are treated with plasma.

82. A fibrosis diagnostic system according to any preceding claim, wherein the plasma comprises oxygen.

83. The fibrosis diagnostic system according to any preceding claim, wherein one or more coatings are applied by: printing, spraying, brushing, injecting, precipitation, polymerization or vapor deposition or any combination thereof.

84. A fibrosis diagnostic system according to any preceding claim, wherein one or more channel surfaces comprise a laminate material.

85. A fibrosis diagnostic system according to any preceding claim, wherein the device housing comprises a transparent material, a translucent material, a colored material, a polarized material, a material that selectively transmits a wavelength of electromagnetic radiation, a material that selectively refracts a wavelength of electromagnetic radiation, a material that selectively reflects a wavelength of electromagnetic radiation, or an opaque material, or any combination thereof.

86. A fibrosis diagnostic system according to any preceding claim, wherein the device housing comprises a window allowing a view of the interior of the device.

87. A fibrosis diagnostic system according to any preceding claim, wherein the view is a view of one or more channels of the plurality of channels.

88. A fibrosis diagnostic system according to any preceding claim, wherein the view comprises a view of the first channel or the target object or both.

89. A fibrosis diagnostic system according to any preceding claim, wherein the device housing comprises a plurality of windows allowing a plurality of views of the interior of the device to be obtained.

90. A fibrosis diagnostic system according to any preceding claim, wherein the target object comprises organic material or inorganic material or both.

91. A fibrosis diagnostic system according to any preceding claim, wherein the target object comprises a tissue or a medical device or both.

92. A fibrosis diagnostic system according to any preceding claim, wherein the target object comprises a spheroid, an organoid, an alginate-based biomaterial, a scaffold-supported cell structure, or a bioprinted structure, or any combination thereof.

93. The fibrosis diagnostic system according to any preceding claim, wherein the target object comprises a tumor cell, a malignant cell or a benign cell or any combination thereof.

94. A fibrosis diagnostic system according to any preceding claim, wherein the target comprises one or more coatings.

95. A fibrosis diagnostic system according to any preceding claim, wherein the one or more coatings of the target are different from the matrix.

96. A fibrosis diagnostic system according to any preceding claim, wherein the one or more coatings of the target differ from one or more coatings of one or more channel surfaces in terms of material composition, thickness or patterning, or a combination thereof.

97. A fibrosis diagnostic system according to any preceding claim, wherein the one or more coatings of the target are identical to the one or more coatings of the one or more channel surfaces in terms of material composition, thickness or patterning or a combination thereof.

98. A fibrosis diagnostic system according to any preceding claim, wherein the target is coated with a plurality of different materials.

99. A fibrosis diagnostic system according to any preceding claim, wherein the target is coated with a plurality of coatings.

100. The fibrosis diagnostic system of any preceding claim, wherein at least two coatings of the plurality of coatings differ in material composition, thickness or patterning, or a combination thereof.

101. A fibrosis diagnostic system according to any preceding claim, wherein the plurality of coatings comprises a first coating adjacent to the target and a second coating covering the first coating, the first coating comprising a first material and the second coating comprising a second material.

102. The fibrosis diagnostic system of any preceding claim, wherein the target is coated prior to placement in the microfluidic device.

103. A fibrosis diagnostic system according to any preceding claim, wherein the matrix comprises a hydrogel.

104. A fibrosis diagnostic system according to any preceding claim, wherein the matrix comprises fibronectin, an extracellular matrix-derived composition, a basement membrane extract, a decellularized extracellular matrix, a dissolved basement membrane preparation, a mixture of extracellular matrix proteins, a tumor-derived composition, matrigel or hyaluronic acid or any combination thereof.

105. The fibrosis diagnostic system according to any preceding claim, wherein the matrix comprises a natural polymer, an artificial polymer or a cross-linked polymer or any combination thereof.

106. The fibrosis diagnostic system according to any preceding claim, wherein the matrix comprises a polysaccharide, a polypeptide, a lipid, a polynucleotide, hyaluronic acid, hyaluronic acid methacrylate, hyaluronic acid diacrylate, fibronectin, fibrinogen, collagen or methacrylate collagen or any combination thereof.

107. A fibrosis diagnostic system according to any preceding claim, wherein the first biological composition comprises fibroblasts or endothelial cells or both.

108. The fibrosis diagnostic system according to any preceding claim, wherein the first biological composition comprises serum, ascorbic acid, retinoic acid, proline or bleomycin or any combination thereof.

109. The fibrosis diagnostic system according to any preceding claim, wherein the first biological composition comprises a biomolecule that promotes fibrosis, a complement protein, a coagulation-related protein, a bacterial cell wall protein, or a lipopolysaccharide (LPS), or any combination thereof.

110. The fibrosis diagnostic system according to any preceding claim, wherein the second biological composition comprises at least one type of biological cell capable of promoting fibrosis.

111. A fibrosis diagnostic system according to any of the preceding claims, wherein the second biological composition comprises factors that promote differentiation of biological cells into biological cell types capable of promoting fibrosis, factors that promote expression of biological molecules that promote fibrosis, or factors that promote expansion of biological cell types capable of promoting fibrosis, or any combination thereof.

112. A fibrosis diagnostic system according to any of the preceding claims, wherein the second biological composition comprises serum, platelets, leukocytes, macrophages, monocytes, peripheral blood mononuclear cells (PBMCs), stem cells, mast cells, dendritic cells, neutrophils, granulocytes, T cells or B cells or any combination thereof.

113. The fibrosis diagnostic system according to any preceding claim, wherein the third biological composition comprises a population of molecules capable of forming a gradient radiating from the third channel into the first channel or into both the first channel and the second channel.

114. The fibrosis diagnostic system according to any preceding claim, wherein the third biological composition comprises a cytokine, a chemokine, a cell nutrient, a transcription factor or a growth factor or any combination thereof.

115. A fibrosis diagnostic system according to any of the preceding claims, wherein the third biological composition comprises CCL-2 / MCP-1, CCL3, CCL11, fibroblast growth factor (FGF), interferon, IFN-γ, interleukin (IL), IL-4, IL-1α, IL-1β, IL-2, IL-5, IL-6, IL-7, IL-9, IL-13, IL-17A, IL-17F, IL-21, IL-22, IL-25, platelet-derived growth factor (PDGF), TGF-α, TGF-β, granulocyte colony stimulating factor (G-CSF), macrophage colony stimulating factor (M-CSF), granulocyte macrophage colony stimulating factor (GM-CSF), prothrombin or thrombin or any combination thereof.

116. A fibrosis diagnostic system according to any preceding claim, wherein the at least one type of molecule of the gradient comprises a cytokine, a chemokine, a cell nutrient, a transcription factor or a growth factor or any combination thereof.

117. A fibrosis diagnostic system according to any preceding claim, wherein the at least one type of molecule of the gradient comprises transforming growth factor beta (TGF-β).

118. The fibrosis diagnostic system according to any preceding claim, wherein the TGF-β comprises TGF-β1.

119. The fibrosis diagnostic system of any preceding claim, wherein the first biological composition, the second biological composition, or the third biological composition, or any combination thereof, comprises labeled cells or labeled biomolecules or both.

120. The fibrosis diagnostic system according to any preceding claim, wherein the first biological composition, the second biological composition, or the third biological composition, or any combination thereof, comprises a compound with known or potential anti-fibrotic efficacy.

121. The fibrosis diagnostic system according to any preceding claim, further comprising the matrix, the first biological composition, the second biological composition, or the third biological composition, or any combination thereof.

122. The fibrosis diagnostic system according to any preceding claim, further comprising the target object.

123. The fibrosis diagnostic system of any preceding claim, further comprising a gradient of at least one molecule across the first channel, the second channel, or the third channel, or any combination thereof.

124. The fibrosis diagnostic system of any preceding claim, wherein one or more components of the matrix, the first biological composition, the second biological composition, or the third biological composition, or any combination thereof, are anchored to a channel surface.

125. The fibrosis diagnostic system of any preceding claim, further comprising a detector comprising a sensor configured to detect fibrosis of the target object or a property indicative of fibrosis of the target object.

126. A fibrosis diagnostic system according to any preceding claim, wherein the detector is configured to receive the microfluidic device.

127. A fibrosis diagnostic system according to any preceding claim, wherein the detector comprises a detector housing comprising a detector interior.

128. The fibrosis diagnostic system of any preceding claim, wherein the detector further comprises a receptacle configured to receive the microfluidic device, the receptacle comprising an opening in the detector housing.

129. A fibrosis diagnostic system according to any preceding claim, further comprising a thermal subsystem.

130. A fibrosis diagnostic system according to any preceding claim, wherein the thermal subsystem comprises a heater or a cooler or both.

131. The fibrosis diagnostic system of any preceding claim, wherein the thermal subsystem is configured to regulate the temperature of the microfluidic device.

132. A fibrosis diagnostic system according to any preceding claim, wherein the detector comprises the thermal subsystem.

133. The fibrosis diagnostic system of any preceding claim, wherein the thermal subsystem is configured to regulate the temperature of the microfluidic device while the microfluidic device is in the receptacle.

134. The fibrosis diagnostic system of any preceding claim, further comprising a fluid subsystem configured for fluid communication with the microfluidic device.

135. The fibrosis diagnostic system of any preceding claim, wherein the fluidic subsystem is configured to load one or more components into the microfluidic device.

136. The fibrosis diagnostic system according to any preceding claim, wherein the fluid subsystem is configured to supply solids, liquids or gases, or any combination thereof, to the microfluidic device.

137. A fibrosis diagnostic system according to any preceding claim, wherein the fluid subsystem is configured to load the target object, the matrix, the first biological composition, the second biological composition, or the third biological composition, or any combination thereof, into the microfluidic device.

138. The fibrosis diagnostic system of any preceding claim, wherein the fluid subsystem is configured to supply one or more nutrients to the microfluidic device.

139. A fibrosis diagnostic system according to any preceding claim, wherein the fluid subsystem is configured to regulate the pressure in the interior of the microfluidic device.

140. The fibrosis diagnostic system of any preceding claim, wherein the fluid subsystem is configured to adjust a gradient across the first channel, the second channel, or the third channel, or any combination thereof.

141. The fibrosis diagnostic system of any preceding claim, wherein the fluidic subsystem is configured to remove one or more components from the microfluidic device.

142. A fibrosis diagnostic system according to any preceding claim, wherein the detector comprises the fluid subsystem.

143. A fibrosis diagnostic system according to any preceding claim, wherein the fluid subsystem is configured for fluid communication with the microfluidic device when the microfluidic device is in the receptacle.

144. A fibrosis diagnostic system according to any preceding claim, wherein the sensor is configured to detect particles or waves or both indicative of fibrosis emitted from the target object or the microfluidic device or both.

145. A fibrosis diagnostic system according to any preceding claim, wherein the sensor is configured to detect electromagnetic radiation.

146. A fibrosis diagnostic system according to any preceding claim, wherein the electromagnetic radiation comprises ultraviolet radiation, visible light or infrared radiation or any combination thereof.

147. A fibrosis diagnostic system according to any preceding claim, wherein the electromagnetic radiation comprises fluorescence or phosphorescence or both.

148. A fibrosis diagnostic system according to any preceding claim, wherein the detector further comprises a particle source or a wave source or both, wherein the particle source or the wave source or both are configured to transmit particles, waves or both to the target object or the microfluidic device or both.

149. A fibrosis diagnostic system according to any preceding claim, further comprising a source of electromagnetic radiation.

150. A fibrosis diagnostic system according to any preceding claim, wherein the detector comprises the electromagnetic radiation source.

151. A fibrosis diagnostic system according to any preceding claim, wherein the electromagnetic radiation source comprises a light emitting diode, a laser, an incandescent light source, an arc light source, a halogen light source, a chemical vapor light source or a fluorescent light source or any combination thereof.

152. A fibrosis diagnostic system according to any preceding claim, wherein the detector further comprises an optical subsystem configured to deliver radiation or affect radiation or both.

153. A fibrosis diagnostic system according to any preceding claim, wherein the optical subsystem comprises one or more lenses, one or more filters, one or more mirrors or one or more diffraction gratings or any combination thereof.

154. A fibrosis diagnostic system according to any preceding claim, wherein the optical subsystem or the sensor or both comprises a microscope or one or more components thereof.

155. A fibrosis diagnostic system according to any preceding claim, wherein the optical subsystem comprises the sensor or the electromagnetic radiation source or both.

156. A fibrosis diagnostic system according to any preceding claim, wherein the sensor is configured to generate an output signal based on an input signal detected by the sensor.

157. A fibrosis diagnostic system according to any preceding claim, wherein the detector further comprises a transmitter, a receiver, a transceiver or any combination thereof, wherein the transmitter, the receiver, the transceiver or any combination thereof is configured to receive information or send information or both.

158. A fibrosis diagnostic system according to any preceding claim, wherein the information comprises the output signal.

159. A fibrosis diagnostic system according to any preceding claim, wherein the information comprises an operating signal for operating the sensor.

160. A fibrosis diagnostic system according to any preceding claim, wherein the detector further comprises a data port.

161. A fibrosis diagnostic system according to any preceding claim, further comprising a processor configured to receive and process information.

162. A fibrosis diagnostic system according to any preceding claim, wherein the processor is configured to control one or more subsystems or components of the fibrosis diagnostic system.

163. The fibrosis diagnostic system of any preceding claim, wherein the processor is configured to control the sensor, the electromagnetic radiation source, the optical subsystem, the thermal subsystem or the fluid subsystem or any combination thereof.

164. A fibrosis diagnostic system according to any preceding claim, wherein the processor is configured to receive the output signal from the sensor, an operational signal from a user input, or both.

165. A fibrosis diagnostic system according to any preceding claim, wherein the detector comprises the processor.

166. The fibrosis diagnostic system of any preceding claim, further comprising a memory configured for communication with the processor.

167. A fibrosis diagnostic system according to any preceding claim, wherein the memory is configured to store a program for controlling the fibrosis diagnostic system or components thereof.

168. A fibrosis diagnostic system according to any preceding claim, wherein the processor comprises the memory.

169. A fibrosis diagnostic system according to any preceding claim, wherein the detector comprises the memory.

170. A fibrosis diagnostic system according to any preceding claim, further comprising a user interface.

171. A fibrosis diagnostic system according to any preceding claim, wherein the user interface comprises a monitor, buttons, keyboard, mouse, light signals, camera, eyepiece lens, speaker or microphone or any combination thereof.

172. A fibrosis diagnostic system according to any preceding claim, wherein the detector comprises the user interface.

173. A fibrosis diagnostic system according to any preceding claim, further comprising a power supply or a power supply interface or both.

174. A fibrosis diagnostic system according to any preceding claim, wherein the detector comprises the power supply or the power supply interface or both.

175. The fibrosis diagnostic system of any preceding claim, further comprising a kit comprising the microfluidic device and one or more reagents for use in the microfluidic device.

176. A fibrosis diagnostic system according to any preceding claim, further comprising a kit comprising: The microfluidic device; and The matrix, the first biological composition, the second biological composition, or the third biological composition, or any combination thereof.

177. A method of analyzing a target subject's susceptibility to fibrosis, the method comprising: placing a target object, a substrate, and a first biological composition in a first channel of a microfluidic device according to any preceding claim; placing a second biological composition in a second channel of the microfluidic device; placing a third biological composition in a third channel of the microfluidic device; as well as The target object or the first channel or both are analyzed to determine the presence or absence of fibrosis associated with the target object.

178. A method according to any preceding claim, wherein the analysis is performed after a predetermined period of time after placing the target object, the matrix, the first biological composition, the second biological composition, or the third biological composition, or any combination thereof, in the microfluidic device.

179. A method according to any preceding claim, wherein the predetermined time period is sufficient to allow a gradient of at least one type of molecule to form.

180. A method according to any preceding claim, wherein the analysis is performed two or more times.

181. A method according to any preceding claim, wherein any two consecutive analyses are separated by a predetermined time period.

182. A method according to any preceding claim, wherein the analysis comprises removing one or more components from the microfluidic device.

183. A method according to any preceding claim, wherein the analysis comprises an immunoassay.

184. A method according to any preceding claim, wherein the analysis comprises the use of antibodies.

185. A method according to any preceding claim, wherein the antibody is labeled.

186. A method according to any preceding claim, wherein the analysis comprises applying a primary antibody and a secondary antibody comprising a detectable label and binding to the primary antibody.

187. The method of any preceding claim, wherein the primary antibody binds to collagen, α-smooth muscle actin, or both.

188. A method according to any preceding claim, wherein the analysis comprises a histological assay.

189. The method of any preceding claim, wherein the analyzing comprises applying picrosirius red dye.

190. The method of any preceding claim, wherein the analyzing comprises visually inspecting the target object or the first channel, or both, for one or more indications of fibrosis associated with the target object.

191. A method according to any preceding claim, wherein the analysis comprises the use of microscopy.

192. A method according to any preceding claim, wherein the analysis comprises the use of one or more spectroscopic techniques.

193. A method according to any of the preceding claims, wherein the analysis comprises nonlinear optical (NLO) multiphoton microscopy, second harmonic generation (SHG) imaging, third harmonic generation (THG) imaging, two-photon excitation fluorescence (TPEF), two-photon excitation fluorescence (TPEF), cell tracker dye imaging, visible light imaging, (ELISA), Griess assay, histopathology, immunostaining, cleared tissue microscopy, or proteomic profiling of homogenized hydrogels, or any combination thereof.

194. A method according to any preceding claim, wherein the analyzing comprises using a detector to detect one or more indicators of fibrosis associated with the target object.

195. The method of any preceding claim, wherein the analysis comprises inserting the microfluidic device into a receptacle.

196. The method of any preceding claim, further comprising preparing the target object, the matrix, the first biological composition, the second biological composition, or the third biological composition, or any combination thereof, from a patient sample.

197. The method of any preceding claim, further comprising implanting the target object into or on a patient.

198. The method of any preceding claim, further comprising implanting a medical device or tissue, or both, into or on a patient, wherein the target object is a representative model of the medical device or tissue, or both.

199. The method of any preceding claim, further comprising removing a medical device or tissue, or both, from a patient, wherein the target object is a representative model of the medical device or tissue, or both.

200. The method of any preceding claim, further comprising administering an anti-fibrotic agent to the patient.

201. The method of any preceding claim, wherein the analysis comprises determining a quantitative measure of fibrosis.

202. The method of any preceding claim, wherein the dose of an anti-fibrotic agent administered to the patient is based on a quantitative measure of the fibrosis.

203. The method of any preceding claim, wherein the anti-fibrotic agent is administered to the patient before, simultaneously with, or after implantation of the target object, tissue associated with the target object, or a medical device associated with the target object, or any combination thereof.

204. The method of any preceding claim, further comprising administering to the patient one or more therapeutic agents in addition to or in place of the anti-fibrotic agent.

205. A method according to any preceding claim, wherein the patient is a mammalian patient or a human patient or both.

206. A method for screening a therapy for efficacy in treating fibrosis, the method comprising: placing a target object, a substrate, and a first biological composition in a first channel of a microfluidic device according to any preceding claim; placing a second biological composition in a second channel of the microfluidic device; placing a third biological composition in a third channel of the microfluidic device; performing the therapy on the microfluidic device; analyzing the target object or the first channel or both to determine the presence, absence or extent of fibrosis associated with the target object; as well as The efficacy of the therapy is determined based on the presence, absence, or difference in the extent of fibrosis in the absence of the therapy.

207. A method according to any preceding claim, wherein the therapy comprises a therapeutic compound and the performing comprises placing the therapeutic compound in the first channel, the second channel, or the third channel, or any combination thereof.

208. The method of any preceding claim, wherein the therapeutic compound comprises a drug compound or a biologic or both.

209. The method of any preceding claim, wherein the therapy comprises modification of the target subject.

210. The method of any preceding claim, wherein the therapy comprises placing a medical device in the first channel, the second channel, or the third channel, or any combination thereof.

211. The method of any preceding claim, wherein the therapy comprises applying radiation to the microfluidic device.

212. The method of any preceding claim, wherein the therapy further comprises treating the patient with a selected therapy.

213. A method for screening a therapy for efficacy in treating cancer, the method comprising: placing a target object, a matrix, and a first biological composition in a first channel of a microfluidic device according to any preceding claim, wherein the target object comprises a tumor cell; placing a second biological composition in a second channel of the microfluidic device; placing a third biological composition in a third channel of the microfluidic device; performing the therapy on the microfluidic device; analyzing the target object or the first channel or both to determine the presence, absence or extent of fibrosis associated with the target object; as well as The efficacy of the therapy is determined based on the difference in the presence, absence, or extent of the cancer in the absence of the therapy.

214. A non-transitory computer readable medium storing a program comprising instructions for performing a method according to any preceding claim.

215. A non-transitory computer readable medium storing a program comprising instructions for controlling a fibrosis diagnostic system according to any preceding claim, or a subsystem thereof, or a component thereof, or any combination thereof.

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