Test chip for analyzing fluids of organ

Abstract

A chip (11) for organ fluid analyzing, includes (a) a first opening (25) formed at an edge of the chip (11) and configured to receive a portion of a droplet of the fluid, (b) a channel (55) extended from the first opening (25) and configured to draw the portion by applying a capillary force to the first opening (25), (c) a chamber (44) configured to contain a flexible blister (66) filled with a staining agent (68), the chamber (44) and the channel (55) have an interface (41) therebetween, which is open and configured to pass at least the fluid between the chamber (44) and the channel (55), and (d) a flexible membrane (77) placed over the flexible blister (66) and covers the chamber (44), and configured to mix between the staining agent (68) and the portion by transferring compressive force to the chamber (44) and puncturing the flexible blister (66).

Description

[0001] TEST CHIP FOR ANALYZING FLUIDS OF ORGAN

[0002] FIELD OF THE INVENTION

[0003] The present invention relates generally to medical devices, and particularly to methods and systems for analyzing fluids of organ using a microfluidic-based test chip.

[0004] BACKGROUND OF THE INVENTION

[0005] Various types of techniques for preparing samples, such as blood, and performing analytical testing thereof (e.g., in laboratories or in the field) are known in the art.

[0006] For example, U.S. Patents 11,865,537 describes a computerized portable diagnostic device configured to perform on-site testing for instantly diagnosing one or more diseases.

[0007] Moreover, U.S. Patent 11,400,447 describes a microfluidic-based test chip for (i) receiving a fluidic sample, and (ii) mixing the fluidic sample with a staining agent using a micromixer.

[0008] SUMMARY OF THE INVENTION

[0009] An embodiment of the present invention that is described herein provides a chip for analyzing a fluid of an organ, the chip includes (a) a first opening, which is (i) formed at an edge of the chip, and (ii) configured to receive at least a portion of at least a droplet of the fluid, (b) a channel, which is extended from the first opening, and is configured to draw the portion by applying a capillary force to the first opening, (c) a chamber, configured to contain a flexible blister filled with a staining agent, the chamber and the channel have an interface therebetween, and the interface is open and configured to pass at least the fluid between the chamber and the channel, and (d) a flexible membrane, which is: (i) placed over the flexible blister and covers the chamber, and (ii) configured to mix between the staining agent and the portion by at least transferring a compressive force to the chamber and puncturing the flexible blister.

[0010] In some embodiments, the chip includes (i) a second opening formed at the edge of the chip besides the first opening, and (ii) a partition, formed at the edge of the chip for separating between the first and second openings, the partition and the first second openings are configured to split the at least a droplet to a first portion in the first opening, and a second portion in the second opening. In other embodiments, the chip includes a first viewing chamber extended from the channel, and a second viewing chamber extended from the second opening, and the partition is extended into the chip for separating between the channel and the second viewing chamber. In yet other embodiments, the chip includes a first breather, a second breather and a third breather that are configured to generate the capillary force in the channel, the first viewing chamber, and the second viewing chamber, respectively. The second breather is coated with a given membrane, and in response to obtaining a mixture between the staining agent and the first portion of the at least a droplet, the given membrane is punctured to obtain the capillary force for drawing the at least a portion of the mixture into the first viewing chamber.

[0011] In some embodiments, the chip includes a stack having (i) a lower plate, (ii) an upper plate, and (iii) a double-sided adhesive tape (DSAT) disposed between and coupled with the upper and lower plates, and at least one of: the chamber, the channel, the first and second openings, and the first and second viewing chambers, are patterned in the DSAT. In other embodiments, the upper plate has one or more bores including at least one of the first, second and third breathers. In yet other embodiments, the upper plate includes a first housing, which is configured to contain at least the chamber and has an upper surface surrounding at least the chamber, and the chip includes a second housing, which is (i) coupled to the flexible membrane, and (ii) configured to seal the chamber.

[0012] In some embodiments, the chip is configured to be inserted into a port of a system having an image sensor, and the first and second viewing chambers are visible to the image sensor for acquiring one or more images of the fluid and the mixture and analyzing properties of the fluid. In other embodiments, the partition is formed at the edge of the chip between the first and second openings, and the partition is configured to cover in the stack, the DSAT, at least part of the upper plate, and at least part of the lower plate. In yet other embodiments, the channel has (a) a first side having a first section interfacing with the chamber and configured to exchange with the chamber at least one of: (i) the fluid, (ii) the staining agent, and (iii) a mixture between the staining agent and the fluid, and (b) a second side, opposite the first side, the second side has a second section interfacing with the first viewing chamber and the second side has a given opening therebetween for conveying the mixture from the channel to the first viewing chamber.

[0013] In some embodiments, the chip includes an additional chamber, and an additional opening between the chamber and the additional chamber, and in response to the transferal of the compressive force to the chamber, the additional chamber is configured to contain at least one of: (i) the fluid, (ii) the staining agent, and (iii) a mixture between the staining agent and the fluid. In other embodiments, in response to releasing the compressive force, the additional opening is configured to transfer, from the additional chamber to at least the chamber, at least part of at least one of: (i) the fluid, (ii) the staining agent, and (iii) the mixture.

[0014] There is additionally provided, in accordance with an embodiment of the present invention, a method for fabricating a chip for analyzing a fluid of an organ, the method includes patterning in a double-sided adhesive tape (DSAT): (i) a first opening at an edge of the DSAT for receiving at least a portion of at least a droplet of the fluid, (ii) a channel, which is extended from the first opening for drawing the portion by applying a capillary force to the portion inserted through the first opening, and (iii) a cavity defining a bottom of a chamber for containing a flexible blister filled with a staining agent. The chamber and the channel have an interface therebetween, and the interface is open for passing at least the fluid between the chamber and the channel. A first side of the DSAT is coupled to a lower plate. An upper plate is fabricated, the upper plate has at least a wall for defining the chamber and for at least partially surrounding the flexible blister. The upper plate is coupled to a second side of the DSAT and the wall and the interface being aligned. A flexible membrane is placed over the flexible blister for covering the chamber and confining the flexible blister.

[0015] In some embodiments, the method includes forming: (i) a second opening at the edge of the DSAT besides the first opening, and (ii) a partition, between the first and second openings, for separating between the first and second openings, wherein the partition and the first second openings are for splitting the at least a droplet to a first portion in the first opening, and a second portion in the second opening. In other embodiments, the method includes forming (i) a first viewing chamber extended from the channel, and (ii) a second viewing chamber extended from the second opening. The partition is extended into the chip for separating between the channel and the second viewing chamber. In yet other embodiments, the method includes (i) forming in the upper plate first, second and third bores of breathers for generating the capillary force in the channel, the first viewing chamber, and the second viewing chamber, respectively, and (ii) coating the second breather with a given membrane.

[0016] In some embodiments, coupling the upper plate to the second side includes aligning the first, second and third bores with the channel, the first viewing chamber, and the second viewing chamber, respectively. In other embodiments, the wall has an upper surface surrounding at least the chamber, and the method includes forming a housing, which is coupled to the flexible membrane, and placing the housing over the upper surface for sealing the chamber.

[0017] In some embodiments, patterning the channel includes forming in the channel: (a) a first side having a first section interfacing with the chamber for exchanging with the chamber at least one of: (i) the fluid, (ii) the staining agent, and (iii) a mixture between the staining agent and the fluid, and (b) a second side, opposite the first side, the second side has a second section interfacing with the first viewing chamber, and forming, in the second section between the channel and the first viewing chamber, a given opening for conveying the mixture from the channel to the first viewing chamber. In other embodiments, the method includes forming an additional chamber, and an additional opening between the chamber and the additional chamber, and in response to applying a compressive force to the flexible membrane and transferring the compressive force from the flexible membrane to the chamber, the flexible blister is punctured and the additional chamber is for containing at least a portion of at least one of: (i) the staining agent, (ii) the fluid, (iii) a mixture between the staining agent and the fluid.

[0018] There is further provided, in accordance with an embodiment of the present invention, a method for preparing a sample for analyzing a fluid of an organ, the method includes receiving a chip including: (a) first and second openings, which are (i) formed at an edge of the chip, (ii) separated by a partition, and (iii) configured to receive at least first and second portions of at least a droplet of the fluid, respectively, (b) a channel, which is extended from the first opening, and is configured to draw the portion by applying a capillary force to the first opening, (c) a chamber, configured to contain a flexible blister filled with a staining agent, the chamber and the channel have an interface therebetween, and the interface is open and configured to pass at least the fluid between the chamber and the channel, (d) a first viewing chamber connected to the channel through a given opening, and a second viewing chamber extended from the second opening into the chip, and (e) a cover of the chamber including (i) a housing, and (ii) a flexible membrane. A flexible blister, which is at least partially confined between the chamber and the cover, is disposed in the chamber. The fluid of the organ is applied to the edge of the chip for splitting the fluid to the first and second portions between the first and second openings, respectively. A compressive force, which is transferred from the flexible membrane to the chamber for puncturing the flexible blister is applied to the flexible membrane, and releasing the compressive force that was applied to the flexible membrane for obtaining a mixture between the first portion of the fluid and the staining agent. In response to obtaining the mixture, a given membrane of a breather connected to the first viewing chamber is punctured for (i) applying a capillary force to the mixture, and (ii) flowing the mixture into the first viewing chamber.

[0019] In some embodiments, obtaining the mixture includes alternately applying and releasing the compressive force multiple times. In other embodiments, the chip includes an additional chamber and an additional opening between the chamber and the additional chamber, and in response to applying the compressive force, at least one of: (i) the fluid, (ii) the staining agent, and (iii) a mixture between the staining agent and the fluid, are contained in the additional chamber. In yet other embodiments, in response to releasing the compressive force, the additional opening transfers, from the additional chamber to at least the chamber, at least part of at least one of: (i) the fluid, (ii) the staining agent, and (iii) the mixture.

[0020] In some embodiments, the chip includes (i) a first breather for applying a first capillary force for drawing the first portion from the first opening into the channel, and (ii) a second breather for applying a second capillary force for drawing the second portion from the second opening into the second viewing chamber, and puncturing the given membrane of a breather includes containing at least a portion of the mixture in the first viewing chamber, and having the second portion contained in the second viewing chamber. In other embodiments, in response to splitting the fluid to the first and second portions between the first and second openings, respectively, inserting the chip into a system for: (i) applying and releasing the compressive force, and (ii) puncturing the given membrane.

[0021] The present invention will be more fully understood from the following detailed description of the embodiments thereof, taken together with the drawings in which:

[0022] BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Fig. 1 is a schematic, top-view illustration of a test chip, in accordance with an embodiment of the present invention;

[0024] Fig. 2 is a schematic, top-view illustration of an upper plate, and a double-sided adhesive tape (DS AT) on a lower plate of the test chip of Fig. 1, in accordance with an embodiment of the present invention;

[0025] Fig. 3 is a schematic, pictorial illustration of a mixer assembly of the test chip of Fig. 1, in accordance with an embodiment of the present invention;

[0026] Fig. 4 is a schematic, sectional view of the mixer assembly of Fig. 3, in accordance with an embodiment of the present invention;

[0027] Fig. 5 is a flow chart that schematically illustrates a method for fabricating the test chip of Fig. 1, in accordance with an embodiment of the present invention; and

[0028] Fig. 6 is a flow chart that schematically illustrates a method for preparing a sample for analysis, using the test chip of Fig. 1, in accordance with an embodiment of the present invention.

[0029] DETAILED DESCRIPTION OF EMBODIMENTS

[0030] OVERVIEW

[0031] Embodiments of the present invention that are described herein provide improved techniques for preparing samples for analyzing fluids of organ, and diagnosing one or more diseases using a microfluidic -based testing device, also referred to herein as a test chip or a chip, for brevity. In some embodiments, the test chip comprises two (e.g., first and second) viewing chambers containing two respective samples for diagnostics. The two samples may be prepared based on a small quantity of a fluid of an organ, in the present example, a single droplet of blood.

[0032] In some embodiments, the test chip comprises first, and second openings formed at the end of the chip, and a partition configured to separate between the first and second openings, so as to split the droplet into first and second portions of blood between the first and second openings, respectively. The chip comprises a channel, which is extended from the first opening, and is configured to convey the first portion from the first opening to (an interface with) a mixer assembly (described below) using a capillary force. The second viewing chamber is shaped as a microfluidic channel, which is extended from the second opening, and is configured to contain the second portion of the blood.

[0033] In some embodiments, the mixer assembly comprises a first chamber (also referred to herein as a mixing chamber) configured to: (i) contain a flexible blister containing a staining agent, and (ii) exchange fluids (e.g., staining agent, blood, and a mixture thereof) with the channel. The mixer assembly may comprise a second chamber and an opening between the first and second chambers. The mixer assembly comprises a cover of the first chamber, the cover comprising a flexible membrane placed on the upper surface of the blister.

[0034] In some embodiments, when preparing the samples, a finger of a subject whose blood is intended to be diagnosed is pricked to extract a droplet of blood, and subsequently, placed over the first and second openings and the partition, so as to split the droplet into first and second portions thereof. When the first portion flows along the channel toward the mixer assembly, a compressive force is applied to the flexible membrane, so as to puncture the blister and start a mixing process between the staining agent and the first portion of blood.

[0035] The mixing process is described in detail in Figs. 1-6 below, but in essence in some embodiments, in response to applying the compressive force, the flexible membrane is inserted into the first chamber, and thereby, reduces the inner volume of the first chamber. As a result, the staining agent flows out of the punctured blister toward the second chamber (which is empty) and the channel (filled with blood) and starts to mix with the blood. Subsequently, releasing the compressive force allows movement of the flexible membrane out of the first chamber, and thereby, increasing the volume of the first chamber to its original volume. The volume increasing results in drawing and mixing the blood and the staining agent within the first chamber. Applying and releasing the compressive force alternately may be carried out several times for obtaining a uniform mixture between the blood and the staining agent. In the present example, the mixture has a blue color. Subsequently, the mixture is conveyed into the first viewing chamber, as will be described below.

[0036] In the context of the present disclosure, the first viewing chamber is also referred to as “a blue channel” for containing the mixture between the staining agent and (the first portion of) blood. And the second viewing chamber is also referred to as “a red channel” for containing only (the second portion of) blood.

[0037] In some embodiments, the chip further comprises three breathers, configured to apply the capillary force for drawing the fluids. More specifically, (a) a first breather is positioned in close proximity to the mixer assembly, and is configured to generate the capillary force for drawing the first portion of blood along the channel, in the present example, from the first opening into the mixer assembly, (b) a second breather is positioned approximately at the edge of the blue channel, and is configured to generate the capillary force for drawing the mixture between (i) the mixer assembly and the channel, and (ii) the blue channel, and (c) a third breather is positioned approximately at the edge of the red channel, and is configured to generate the capillary force for drawing the second portion of blood from the second opening into the red channel.

[0038] In some embodiments, the second breather is coated with a membrane. After obtaining the mixture (of blood and staining agent), the second breather is punctured so as to apply the capillary force and draw the mixture into the blue channel. The sample preparation is concluded when the red and blue channels are filled with the blood and the mixture, respectively.

[0039] In some embodiments, after obtaining the subject’s blood droplet, the test chip is inserted into a system that automatically (i) performs the sample preparation process described above, and subsequently, (ii) acquires one or more images of the fluids in the red and blue channels, and processes the images for diagnosing whether or not the blood contains indications to one or more diseases.

[0040] The disclosed test chip enable preparation of two or more samples from a single fluid of an organ intended to be tested. In the embodiments described above, the samples comprise (i) blood, and (ii) the mixture between blood and the staining agent, and the system may perform the diagnostics by comparing between processed images acquired from the red and blue channels, or using any other diagnostic technique. In other embodiments, the chip may comprise two or more sets of staining agents, mixer assemblies, viewing chambers etc., so as to increase variety of diseases that could be tested using the chip and the diagnostic system. SYSTEM DESCRIPTION

[0041] Fig. 1 is a schematic, top-view illustration of a test chip 11, in accordance with an embodiment of the present invention. Test chip 11 comprises a device containing one or more samples produced based on a fluid of an organ. The test chip 11 is configured to be inserted into a system, so as to analyze the sample as will be described below. It is noted that Fig. 1 is a general overview of test chip 11, and the components of test chip 11 are described in more detail in Figs. 2-4 below.

[0042] In some embodiments, test chip 11, also referred to herein as a chip 11, for brevity, is configured to receive a fluidic sample of a subject. In the present example, the fluidic sample may comprise any suitable fluid of a human body, such as but not limited to tissue, blood, urine, sputum and the like. In other embodiments, the fluidic sample may comprise any suitable fluid of an organ other than fluid of a human body. In the description below, the fluid sample comprises a droplet of blood (or multiple droplets of blood) intended to be tested using chip 11 , and in other embodiments, the structure and functionality of chip 11 may be used, mutatis mutandis, for testing the other sorts of fluid samples, such as the fluids described above.

[0043] In some embodiments, test chip 11 comprises openings 25 and 26, formed at an edge of chip 11 configured to inlet first and second portions of the droplet of blood, respectively, into respective microfluidic channels of chip 11 that are described in detail below.

[0044] In some embodiments, chip 11 comprises a partition 27 separating between openings 25 and 26 and configured to split the blood droplet into the aforementioned first and second portions that are intended to be inserted through openings 25 and 26, respectively. Moreover, partition 27 is configured to allow the blood to flow through both openings at the same time in a parallel flow without having mutual interferences. As a result, the blood composition within openings 25 and 26 remains substantially similar.

[0045] In some embodiments, chip 11 further comprises (i) a mixer assembly 33 having a cover 75, which comprises a housing 76, and a flexible membrane 77, and (ii) a channel 55 extending between opening 25 and mixer assembly 33, for conveying the blood therebetween. The structure and functionality of mixer assembly 33 and channel 55 are described in more detail in Figs. 2-4 below. In the present example, membrane 77 of cover 75 is formed, for example, by silicone injection (or any other suitable process of silicone or another suitable material) and has an elongation between about 120% and 200%, and a hardness of SHORE 40A.

[0046] In the present example, during the sample preparation, a finger of a patient (not shown) whose blood is pricked to extract a droplet of blood intended to be diagnosed, and the blooded finger is placed over openings 25 and 26, so that partition 27 splits the first and second portions of the blood droplet to openings 25 and 26, respectively. The first portion is conveyed by channel 55 between opening 25 and mixer assembly 31. Test chip 31 comprises a breather 28, which is configured to enable capillary force applied for drawing the first portion into mixer assembly 33. In the present example, breather 28 is partially covered by housing 76 of cover 75.

[0047] In some embodiments, test chip 11 comprises chambers 44 and 46 having an opening therebetween, as will be shown and described in detail in Fig. 2 below. Chamber 44, also referred to herein as a mixing chamber, is configured to contain a blister (shown and described in detail in Figs. 3 and 4 below).

[0048] In some embodiments, the blister has a flexible member configured to contain a staining agent, also referred to herein as stain. Mixer assembly 33 is configured to generate a mixture between the stain and the blood received from opening 25. More specifically, a compressive force is applied to flexible membrane 77, typically by a diagnostic system of the blood, but could done be manually or using another suitable apparatus.

[0049] In some embodiments, in response to the compressive force applied to membrane 77, the blister is configured to rupture, and subsequently, the stain and the blood are being mixed by alternately applying and releasing the compressive force to membrane 77, as will be described in more detail in Figs. 2-4 below.

[0050] In some embodiments, test chip 11 comprises a viewing chamber implemented in a channel 35, and a breather 45 located at the end of channel 35. Breather 45 has a membrane (not shown) and after generating the mixture between the stain and the blood, the membrane of breather 45 is being punctured so as to generate capillary force that draws the mixture to flow from (mixer assembly 33 and) channel 55 into channel 35 and toward breather 45, e.g., along an X-axis of chip 11.

[0051] In some embodiments, test chip 11 comprises an additional viewing chamber implemented in a channel 36, which is: (i) extended from opening 26, and (ii) configured to generate capillary force that draws the second portion of the blood droplet to flow from opening 26 into channel 36 toward a breather 48.

[0052] In some embodiments, test chip 11 is configured to be inserted into a system for testing and diagnosing various types of diseases in body fluids, for example, by performing imaging and visual inspection of body fluids such as blood and a mixture between the body fluid and a staining agent, as described above. Such systems are described, for example, in PCT International Publication WO 2023 / 079435, and in U.S. Patents 11,865,537, and 11,400,447, whose disclosures are all incorporated herein by reference. In the present example, the system comprises a hand-held computerized microscope, which is carried in the field e.g., by a user, and is configured to perform on-site testing.

[0053] In some embodiments, the system (not shown) has one or more ports configured for receiving chip 11 or any other suitable type of a disposable sample having blood sample for analysis. The system comprises an imaging device, such as a computerized microscope configured to acquire images of the blood (or any other body fluids) and the mixture described above, in channels 36 and 35, respectively. Moreover, the system may comprise (i) a movable stage configured to move chip 11 along one or more axes for scanning and acquiring the images, and (ii) a processor, configured to analyze the acquired images for detecting whether or not the images are indicative of one or more diseases in the subject whose blood has been tested by the system.

[0054] Typically, the processor of the system comprises a general-purpose computer, which is programmed in software to carry out the functions described herein. The software may be downloaded to the computer in electronic form, over a network, for example, or it may, alternatively or additionally, be provided and / or stored on non-transitory tangible media, such as magnetic, optical, or electronic memory.

[0055] In some embodiments, the system is configured to: (a) perform the mixing between the blood and the staining agent by alternately (i) applying the compressive force (e.g., pressure) to membrane 77, and (ii) releasing the pressure from membrane 77, for obtaining the mixture, and subsequently (b) puncturing at least breather 45 for drawing the mixture into chamber 35 as described above.

[0056] In some embodiments, test chip 11 comprises a stack of an upper plate 22, a lower plate, and a double-sided adhesive tape (shown in Fig. 2 below), which is disposed between the upper and lower plates. The upper and lower plates may comprise a suitable polymer such as but not limited to polymethylmetacrylate (PMMA), cyclo-olefin-copolymer (COC), cycloolefinpolymer (COP), polycarbonate (PC), and polystyrene (PS). The polymer has a typical transparency larger than about 92% to light in wavelengths of the visible spectrum (e.g., between about 380 nm and 750 nm. Such plates are implemented, for example, using the ZEONEX 5000 product, or any other suitable product from the ZEONEX family of products, supplied by ZEON CORPORATION (1-6-2 Marunouchi, Chiyoda-Ku, Tokyo, 100-8246, Japan). The polymer is being formed and shaped using plastic molding, and the plastic molds may require some protrusions and / or intrusions 31a and 31b formed at least in upper plate 22 during the plastic injection molding process. In the present example, test chip 11 has a size between about (i) 40 mm and 80 mm along the X-axis, (ii) 18 mm and 35 mm along the Y-axis, and (iii) a thickness between about 4 mm and 10 mm along the Z-axis. For example, the lower plate has a thickness of about 0.175 mm, and the double-sided adhesive tape has a thickness of about 0.1 mm, and the thickness of upper plate 22 is between about 2 mm and 8 mm. Moreover, mixer assembly 33 has a diameter between about 10 mm and 20 mm, and channels 35 and 36 and openings 25 and 26 have a cross section size between about 1 mm and 3 mm.

[0057] Fig. 2 is a schematic, top-view illustration of upper plate 22, and a double-sided adhesive tape (DS AT) 15 disposed on a lower plate 24 of test chip 11, in accordance with an embodiment of the present invention. DSAT 15 is stacked on plate 24, and plate 22 is placed on DSAT 15, thus plates 22 and 24 are also referred to herein as the upper and lower plates of test chip 11, respectively.

[0058] In some embodiments, may comprise a pressure-sensitive adhesive that may be based on at least one of: (i) natural rubber, (ii) synthetic rubber, (iii) acrylic, and (iv) silicone. In the TM present example, DSAT 15 comprises 3M Double Coated Tape 9629B, supplied by 3M Corporate (St. Paul, MN 55144-1000).

[0059] In the example of Fig. 2, cover 75 and blister 66 are removed from the top-view illustration of plate 22 for the sake of presentation, and are depicted in detail in Figs. 3 and 4 below.

[0060] In some embodiments, as described in Fig. 1 above, the blood is inserted into opening 25 and flows along channel 55. In the present example, channel 55 has a U-shape and surrounding at least part of chamber 44, but in other implementations, channel 55 may have any other suitable shape such as but not limited to a round (e.g., “O”) shape.

[0061] In some embodiments, chambers 44 and 46 and channel 55, openings 25, 26, and 49, and channels 35 and 36, are patterned through (i.e., removing corresponding sections of) DSAT 15, which is subsequently stacked on and glued to the surface of plate 24. Plate 24 is flat, so that the above patterning in DSAT 15 enable the flowing of: (i) the first portion of the blood droplet from opening 25, through channel 55, chambers 46 and 44, and opening 49, into channel 35 that serves as the viewing chamber of the mixture, and (ii) the second portion of the blood droplet from opening 26 into channel 36 that serves as the viewing chamber of the blood. As described above, the system is configured to move and scan test chip 11 relative to the microscope for acquiring images of the fluid in channels 35 and 36.

[0062] In some embodiments, channel 55 is confined between plates 22 and 24 along the Z- axis. In the XY plane, channel 55 has a U-shape with an inner arc-shaped side 41 and an outer arc-shaped side 42 extended between opening 25 and chamber 46, thereby surrounding at least a portion of chamber 44. Reference is now made to the top view of plate 24 and DSAT 15. The inner arc-shaped side 41 of channel 55 is marked with a dashed line, and the outer arc-shaped side 42 of channel 55 is marked with a solid line. A sectional view of channel 55 is illustrated and described in detail in Fig. 4 below.

[0063] In some embodiments, inner side 41 of channel 55 that interfaces with chamber 44 is marked with a dashed line to indicate that side 41 is open (e.g., there is an opening between channel 55 and chamber 44 along the dashed line), so that, in the present example embodiments, this opening allows fluids to flow through inner arc-shaped side 41, and thereby, configured to facilitate exchanging of the fluids between channel 55 and chamber 44. In such embodiments, side 41 functions as an interface between chamber 44 and channel 55. The outer side 42 of channel 55 has a small section, in the present example an opening 49. In response to the puncturing of breather 45, opening 49 enables the flow of the mixture (as described in Fig. 1 above) between channel 55 and channel 35. The remaining sections of outer side 42 of channel 55 has the full stack of plates 22 and 24 and DSAT 15 therebetween, thus, the remaining outer side 42 of channel 55 is configured to block the passage of fluids. It is noted that breather 28 is open, and channel 55 is extended between opening 25 and the opening of breather 28 (breathers 28, 45, and 48 are shown in the top view of plate 22).

[0064] In such embodiments, the capillary force is applied to the first portion of the blood along microfluidic channel 55, so as to convey the blood from opening 25 toward mixer assembly 33. Moreover, when breather 45 is blocked by the membrane (before obtaining the mixture as described above), the blood flowing in channel 55 is not passing through opening 49 into channel 35. After obtaining the mixture between the blood and the staining agent (as described in Fig. 1 above), the membrane coating breather 45 is punctured (e.g., by the system, or using any other technique), and the capillary force is applied to the mixture for drawing the mixture through opening 49 and along channel 35.

[0065] In some embodiments, breather 48 is open, so that microfluidic channel 36 extends between opening 26 and the opening of breather 48. Thus, in response to inserting the second portion of the blood droplet into opening 26, the capillary force is applied to the blood, and resulting in the drawing of the blood along channel 36.

[0066] In the present example, the mixture has a blue color, and the blood has a red color. In the context of the present disclosure, the first viewing chamber, which is implemented in microfluidic channel 35, is also referred to as “a blue channel” for containing the mixture between the staining agent and (the first portion of) blood. Moreover, the second viewing chamber, which is implemented in microfluidic channel 36, is also referred to as “a red channel” for containing only (the second portion of) blood.

[0067] In some embodiments, chamber 46 is partially surrounded by an arc-shaped line 43 patterned in DSAT 15.

[0068] Reference is now made to the top view of plate 22. In some embodiments, chip 11 comprises a structure, which is formed over the outer surface of plate 22 and is defined by: (i) a wall 51 surrounding and defining chamber 44, (ii) a wall 50 is formed over arc-shaped line 43 for partially surrounding and defining chamber 46, and (iii) an upper surface 52.

[0069] In some embodiments, wall 51 of the structure has a tapered profile, (as also shown in a sectional view in Fig. 4 below), and is configured to confine blister 66 within chamber 44. In some embodiments, in response to alternately (i) applying the compressive force to membrane 77, and (ii) releasing the compressive force off membrane 77, the blood flowing in channel 55 and the staining agent (that was contained in blister 66 and has been flowing in chambers 44 and 46 responsively to puncturing blister 66) are mixing in chambers 44 and 46 and in channel 55, as described in Fig. 1 above.

[0070] In some embodiments, chip 11 has an opening 47 between chambers 44 and 46. In response to the reduced volume of chamber 44, at least a portion of the staining agent (that was contained in the punctured blister 66) is being pushed to flow out of chamber 44, through opening 47, and into chamber 46. Moreover, in response to releasing the compressive force off membrane 77, the volume of chamber 44 increases, at least a portion of the staining agent flows through opening 47 back into chamber 44 and mixes with the blood flowing from channel 55 into chamber 44.

[0071] In some embodiments, the system is configured to alternately (i) press on membrane 77, and (ii) release the pressure off membrane 77, several times so as to generate a uniform mixing between the staining agent and the first portion of the blood droplet that is inserted through opening 25 and channel 55 as described above. As such, chamber 44 functions as a mixing chamber between the staining agent and the fluid (e.g., blood) intended to be analyzed by the system, as described above. The number of pressing and releasing cycles depends on the properties (e.g., viscosity) of the staining agent and the fluid sample, the affinity between the staining agent and the fluid sample, and the analysis application. For example, between about 20 cycles and 30 cycles may be required to obtain a uniform mixture between the staining agent and the blood.

[0072] In some embodiments, after obtaining the required level of mixture, the system is configured to puncture breather 45 so as to generate the capillary force that draws the mixture of staining agent and blood to flow from chamber 44, along the X-axis of chip 11, and into channel 35. In some embodiments, in response to puncturing breather 45, the capillary force draws the mixture from chamber 44, though opening 49, and into channel 35, as described above. As shown by (virtual) dashed lines 13 and 14, breathers 48 and 45 are located at the edge of channels 36 and 35, respectively, so as to enable the generating of the capillary force, as described above.

[0073] Fig. 3 is a schematic, pictorial illustration of mixer assembly 33, in accordance with an embodiment of the present invention.

[0074] In some embodiments, cover 75 comprises housing 76, which is placed over upper surface 52, and snugly fits (i) in chamber 44, and (ii) over blister 66 that contains staining agent 68. Cover 75 further comprises membrane 77, which is flexible and is coupled to housing 76. In some embodiments, the application of the compressive force to membrane 77 reduces the volume of chamber 44, and the releasing of the compressive force off membrane 77, increased the volume of chamber 44 to its original volume (i.e., before applying the compressive force to membrane 77). Cover 75 is placed over the outer surface of blister 66, as will be shown and depicted in Fig. 4 below.

[0075] Fig. 4 is a schematic, sectional view of mixer assembly 33, in accordance with an embodiment of the present invention.

[0076] In some embodiments, membrane 77 is coupled with housing 76 using elastic U-shaped connectors 53 that enable the flexibility (i) while pressing on membrane 77, and subsequently (ii) when releasing the pressure off membrane 77. When housing 76 is placed over surface 52, the elastic U-shaped connectors 53 fit in wall 51, so as to provide membrane 77 with mechanical support. Membrane 77 is placed over blister 66, so that in response to applying the compressive force to membrane 77, the membrane 77 is distorted into chamber 44, so that blister 66 is crushed and subsequently punctured or explodes. In response to the puncturing of blister 66, the staining agent 68 scatters within the volume of chamber 44, and flows into chamber 46 as shown and described in Fig. 2 above. As described above, the volume of chamber 44 increases and decreases alternately to generate the mixture of the blood and staining agent 68. The alternating compressive force applied to and released off membrane 77 repeats until obtaining the desired mixture between the blood and the staining agent 68. In the present example, blister 66 has a dome shape facing membrane 77 and a base, which is typically wider than the dome in XY plane and facing wall 50. In other embodiments, blister 66 may have any other suitable shape, such as a trapezoid-like shape whose small base facing membrane 77 and large base facing wall 50. In alternative embodiments, blister 66 may be disposed in chamber 44 in a flipped orientation (e.g., up-side-down), such that the dome or small base is facing wall 50 and large base is facing membrane 77.

[0077] Reference is now made to an inset 57 showing a sectional view of channel 55. In some embodiments, DSAT 15 is disposed between upper plate 22 and lower plate 24, and channel 55 is patterned in DSAT 15 and is defined, in the XY plane, between (i) outer arc-shaped side 42 interfacing between channel 55 and DSAT 15, and (ii) inner arc-shaped side 41 that is open between channel 55 and chamber 44, as shown and described in the top view of plate 24 and DSAT 15 of Fig. 2 above.

[0078] In the present example, wall 51 is formed on upper plate 22 and has a tapered profile as shown also in the general view of Fig. 4. It is noted that the edge of wall 51 is aligned with inner arc-shaped side 41 along the Z-axis.

[0079] Fig. 5 is a flow chart that schematically illustrates a method for fabricating test chip 11, in accordance with an embodiment of the present invention.

[0080] The method begins at a patterning step 100 with patterning in DSAT 15 (i) openings 25 and 26, (ii) chambers 44 and 46 and opening 47, (iii) channels 35 and 36, and (iv) opening 49, and glueing DSAT 15 on the surface of plate 24, as described in detail in Fig. 2 above.

[0081] At a mixer and breather formation step 102, mixer assembly 33 is formed over the surface of plate 22 by forming (i) wall 50 of chamber 46 and wall 51 of chamber 44, on plate 22, and (ii) bores (e.g., drills) of breathers 28, 45 and 48, as described in detail in Fig. 2 above.

[0082] At an upper plate aligning and stacking step 104, upper plate 22 is stacked over DSAT 15 and lower plate 24. Moreover, the bores of breather 28 is aligned with chamber 46, and the bores of breathers 48 and 45 are aligned with the edge of channels 36 and 35, respectively, as described in detail in Fig. 2 above. It is noted that DSAT 15 has glue on both sides, so that after the alignment, upper plate 22 is stacked on and glued to DSAT 15.

[0083] At a partitioning step 106, partition 27 is formed for separating between openings 25 and 26, as described in detail in Fig. 2 above.

[0084] At a mixer covering step 108 that concludes the method, blister 66 is disposed within chamber 44, and subsequently, cover 75 is fitted within chamber 44 so that housing 76 is placed over surface 52, and membrane 77 is placed over blister 66, as described in detail in Figs. 3 and 4 above. In other embodiments, blister 66 may not be inserted during the fabrication, but only during the utilization of test chip 11 for preparing a sample for testing in the system, as will be described in Fig. 6 below. In such embodiments, only cover 75 is fitted within chamber 44 and housing 76 is placed over surface 52. It is noted that the order of steps may be different than that described above, and that some of the above steps may be carried out simultaneously.

[0085] Fig. 6 is a flow chart that schematically illustrates a method for preparing a sample for analysis, using test chip 11, in accordance with an embodiment of the present invention.

[0086] The method begins at a blister placement step 200, with lifting cover 75, placing blister 66 (that contains staining agent 68) over chamber 44, and placing cover 75 over blister 66, as shown and described in detail in Fig. 3 above.

[0087] At a fluid sample insertion step 202, the finger of the subject is pricked to extract the droplet of blood, which is inserted into chip 11 through openings 25 and 26, as described in detail in Figs. 1 and 2 above. It is noted that the droplet of blood splits between openings 25 and 26, and due to the capillary force, (i) the first portion of the blood droplet flows through channel 55 toward mixer assembly 33, and (ii) the second portion of the blood droplet flows through channel 36, also referred to as the second viewing chamber and the red channel, as described in detail in Figs. 1 and 2 above.

[0088] At a mixing step 204, a repetitive alternating process comprising: (i) pressing on membrane 77 for puncturing / exploding blister 66 and flowing staining agent 68 into chambers 44 and 46 and channel 55, and subsequently, (ii) releasing the pressure off membrane 77, is carried out several times (e.g., about 30 times) for mixing between staining agent 68 and the first portion of the blood droplet inserted through opening 25. It is noted that step 204 may be carried out (a) by the system after inserting test chip 11 into a port of the system, as described in detail in Fig. 1 above, or (b) manually (using an operator’s finger), or using any suitable apparatus configured to alternately press on membrane 77 and release the pressure off membrane 77.

[0089] At a mixture flowing step 206 that concludes the method, after obtaining the required mixture (between the staining agent 68 and the blood) in chamber 44 and channel 55, puncturing breather 45 for applying the capillary force to the mixture, in order to draw the mixture into channel 35, which is also referred to as the first viewing chamber, and the blue channel, as described in detail in Figs. 1 and 2 above.

[0090] In some embodiments, after channel 35 is filled with the mixture, and channel 36 is filled with the blood, the channels 35 and 36 serve as viewing chambers for the microscope (or any other imaging device) to diagnose one or more diseases in the blood intended to be tested, as described in detail in Fig. 1 above. It is noted that the method of Fig. 6 may be applied, mutatis mutandis, to test chip 11 for preparing the samples for analysis using the system described in detail in Fig. 1 above, as well as in the aforementioned U.S. Patents 11,865,537, and 11,400,447, whose disclosures are all incorporated herein by reference.

[0091] Although the embodiments described herein mainly address test chip for analyzing blood, the methods and systems described herein can also be used in other applications. It will thus be appreciated that the embodiments described above are cited by way of example, and that the present invention is not limited to what has been particularly shown and described hereinabove. Rather, the scope of the present invention includes both combinations and sub-combinations of the various features described hereinabove, as well as variations and modifications thereof which would occur to persons skilled in the art upon reading the foregoing description and which are not disclosed in the prior art. Documents incorporated by reference in the present patent application are to be considered an integral part of the application except that to the extent any terms are defined in these incorporated documents in a manner that conflicts with the definitions made explicitly or implicitly in the present specification, only the definitions in the present specification should be considered.

Claims

CLAIMS1. A chip for analyzing a fluid of an organ, the chip comprising: a first opening, which is (i) formed at an edge of the chip, and (ii) configured to receive at least a portion of at least a droplet of the fluid; a channel, which is extended from the first opening, and is configured to draw the portion by applying a capillary force to the first opening; a chamber, configured to contain a flexible blister filled with a staining agent, wherein the chamber and the channel have an interface therebetween, and wherein the interface is open and configured to pass at least the fluid between the chamber and the channel; and a flexible membrane, which is: (i) placed over the flexible blister and covers the chamber, and (ii) configured to mix between the staining agent and the portion by at least transferring a compressive force to the chamber and puncturing the flexible blister.

2. The chip according to claim 1, and comprising (i) a second opening formed at the edge of the chip besides the first opening, and (ii) a partition, formed at the edge of the chip for separating between the first and second openings, wherein the partition and the first second openings are configured to split the at least a droplet to a first portion in the first opening, and a second portion in the second opening.

3. The chip according to claim 2, and comprising a first viewing chamber extended from the channel, and a second viewing chamber extended from the second opening, wherein the partition is extended into the chip for separating between the channel and the second viewing chamber.

4. The chip according to claim 3, and comprising a first breather, a second breather and a third breather that are configured to generate the capillary force in the channel, the first viewing chamber, and the second viewing chamber, respectively, wherein the second breather is coated with a given membrane, and wherein, in response to obtaining a mixture between the staining agent and the first portion of the at least a droplet, the given membrane is punctured to obtain the capillary force for drawing the at least a portion of the mixture into the first viewing chamber.

5. The chip according to claim 4, and comprising a stack having (i) a lower plate, (ii) an upper plate, and (iii) a double-sided adhesive tape (DSAT) disposed between and coupled with the upper and lower plates, wherein at least one of: the chamber, the channel, the first and second openings, and the first and second viewing chambers, is patterned in the DSAT.

6. The chip according to claim 5, wherein the upper plate has one or more bores comprising at least one of the first, second and third breathers.

7. The chip according to claim 5, wherein the upper plate comprises a first housing, which is configured to contain at least the chamber and has an upper surface surrounding at least the chamber, and comprising a second housing, which is (i) coupled to the flexible membrane, and (ii) configured to seal the chamber.

8. The chip according to claim 5, wherein the chip is configured to be inserted into a port of a system having an image sensor, and wherein the first and second viewing chambers are visible to the image sensor for acquiring one or more images of the fluid and the mixture and analyzing properties of the fluid.

9. The chip according to claim 5, wherein the partition is formed at the edge of the chip between the first and second openings, and the partition is configured to cover in the stack, the DSAT, at least part of the upper plate, and at least part of the lower plate.

10. The chip according to any of claims 1-9, wherein the channel has (a) a first side having a first section interfacing with the chamber and configured to exchange with the chamber at least one of: (i) the fluid, (ii) the staining agent, and (iii) a mixture between the staining agent and the fluid, and (b) a second side, opposite the first side, the second side has a second section interfacing with the first viewing chamber and the second side has a given opening therebetween for conveying the mixture from the channel to the first viewing chamber.

11. The chip according to any of claims 1-9, and comprising an additional chamber, and an additional opening between the chamber and the additional chamber, wherein in response to the transferal of the compressive force to the chamber, the additional chamber is configured to contain at least one of: (i) the fluid, (ii) the staining agent, and (iii) a mixture between the staining agent and the fluid.

12. The chip according to claim 11, wherein in response to releasing the compressive force, the additional opening is configured to transfer, from the additional chamber to at least the chamber, at least part of at least one of: (i) the fluid, (ii) the staining agent, and (iii) the mixture between the staining agent and the fluid.

13. A method for fabricating a chip for analyzing a fluid of an organ, the method comprising: patterning in a double-sided adhesive tape (DSAT): (i) a first opening at an edge of theDSAT for receiving at least a portion of at least a droplet of the fluid, (ii) a channel, which is extended from the first opening for drawing the portion by applying a capillary force to theportion inserted through the first opening, and (iii) a cavity defining a bottom of a chamber for containing a flexible blister filled with a staining agent, wherein the chamber and the channel have an interface therebetween, and wherein the interface is open for passing at least the fluid between the chamber and the channel; coupling a first side of the DSAT to a lower plate; fabricating an upper plate having at least a wall for defining the chamber and for at least partially surrounding the flexible blister; coupling the upper plate to a second side of the DSAT and aligning between the wall and the interface; and placing a flexible membrane over the flexible blister for covering the chamber and confining the flexible blister.

14. The method according to claim 13, and comprising forming: (i) a second opening at the edge of the DSAT besides the first opening, and (ii) a partition, between the first and second openings, for separating between the first and second openings, wherein the partition and the first second openings are for splitting the at least a droplet to a first portion in the first opening, and a second portion in the second opening.

15. The method according to claim 14, and comprising forming (i) a first viewing chamber extended from the channel, and (ii) a second viewing chamber extended from the second opening, wherein the partition is extended into the chip for separating between the channel and the second viewing chamber.

16. The method according to claim 15, and comprising (i) forming in the upper plate first, second and third bores of breathers for generating the capillary force in the channel, the first viewing chamber, and the second viewing chamber, respectively, and (ii) coating the second breather with a given membrane.

17. The method according to claim 16, wherein coupling the upper plate to the second side comprises aligning the first, second and third bores with the channel, the first viewing chamber, and the second viewing chamber, respectively.

18. The method according to claim 13, wherein the wall has an upper surface surrounding at least the chamber, and comprising forming a housing, which is coupled to the flexible membrane, and placing the housing over the upper surface for sealing the chamber.

19. The method according to any of claims 15-18, wherein patterning the channel comprises forming in the channel: (a) a first side having a first section interfacing with the chamber forexchanging with the chamber at least one of: (i) the fluid, (ii) the staining agent, and (iii) a mixture between the staining agent and the fluid, and (b) a second side, opposite the first side, the second side has a second section interfacing with the first viewing chamber, and forming, in the second section between the channel and the first viewing chamber, a given opening for conveying the mixture from the channel to the first viewing chamber.

20. The method according to any of claims 15-18, and comprising forming an additional chamber, and an additional opening between the chamber and the additional chamber, wherein in response to applying a compressive force to the flexible membrane and transferring the compressive force from the flexible membrane to the chamber, the flexible blister is punctured and the additional chamber is for containing at least a portion of at least one of: (i) the staining agent, (ii) the fluid, (iii) a mixture between the staining agent and the fluid.

21. A method for preparing a sample for analyzing a fluid of an organ, the method comprising: receiving a chip comprising: first and second openings, which are (i) formed at an edge of the chip, (ii) separated by a partition, and (iii) configured to receive at least first and second portions of at least a droplet of the fluid, respectively; a channel, which is extended from the first opening, and is configured to draw the portion by applying a capillary force to the first opening; a chamber, configured to contain a flexible blister filled with a staining agent, wherein the chamber and the channel have an interface therebetween, and wherein the interface is open and configured to pass at least the fluid between the chamber and the channel; a first viewing chamber connected to the channel through a given opening, and a second viewing chamber extended from the second opening into the chip; and a cover of the chamber comprising (i) a housing, and (ii) a flexible membrane; disposing, in the chamber, a flexible blister at least partially confined between the chamber and the cover; applying the fluid of the organ to the edge of the chip for splitting the fluid to the first and second portions between the first and second openings, respectively; applying to the flexible membrane (i) a compressive force, which is transferred from the flexible membrane to the chamber for puncturing the flexible blister, and (ii) releasing the compressive force for obtaining a mixture between the first portion of the fluid and the staining agent; andin response to obtaining the mixture, puncturing a given membrane of a breather connected to the first viewing chamber for (i) applying a capillary force to the mixture and (ii) flowing the mixture into the first viewing chamber.

22. The method according to claim 21, wherein obtaining the mixture comprises alternately applying and releasing the compressive force multiple times.

23. The method according to claim 21, wherein the chip comprises an additional chamber, and an additional opening between the chamber and the additional chamber, wherein in response to applying the compressive force, at least one of: (i) the fluid, (ii) the staining agent, and (iii) a mixture between the staining agent and the fluid, are contained in the additional chamber.

24. The method according to claim 23, wherein in response to releasing the compressive force, the additional opening transfers, from the additional chamber to at least the chamber, at least part of at least one of: (i) the fluid, (ii) the staining agent, and (iii) the mixture between the staining agent and the fluid.

25. The method according to claim 21, wherein the chip comprises (i) a first breather for applying a first capillary force for drawing the first portion from the first opening into the channel, and (ii) a second breather for applying a second capillary force for drawing the second portion from the second opening into the second viewing chamber, and wherein puncturing the given membrane of a breather comprises containing at least a portion of the mixture in the first viewing chamber, and having the second portion contained in the second viewing chamber.

26. The method according to any of claims 21-25, wherein, in response to splitting the fluid to the first and second portions between the first and second openings, respectively, inserting the chip into a system for: (i) applying and releasing the compressive force, and (ii) puncturing the given membrane.

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