Microarray, hybridization encapsulation unit, and related method

Abstract

Microarrays, hybridization seals, and related methods. The device includes a substrate containing a plurality of probes and a hybridization seal. The hybridization seal includes an evaporation barrier and a layer including walls, the walls forming a grid pattern and defining a plurality of sample chambers for receiving fluid. The layer includes a first side removably coupled to the substrate and a second side coupled to the evaporation barrier. The evaporation barrier includes a barrier section that covers the probes and includes one or more slits that allow the barrier section to have a convex or concave profile depending on the amount of fluid in the corresponding sample chamber.

Description

Technical Field

[0001] Related Application Section This application claims the benefit and priority of U.S. Provisional Patent Application No. 63 / 051,855, filed on July 14, 2020, the content of which is hereby incorporated by reference in its entirety for all purposes. Background Art

[0002] A microarray can be imaged by an imaging system to detect the presence of a particular target of interest. Before imaging the substrate, the substrate can undergo a hybridization process.

Summary of the Invention

[0003] By providing the microarrays, hybridization seals, and related methods described herein, the disadvantages of the prior art can be overcome and the benefits described hereinafter in the present disclosure can be achieved. Various implementations of the apparatus and method are described below, and the apparatus and method can overcome these disadvantages and achieve the benefits described herein in any combination (on the premise that these combinations are not contradictory) that includes and excludes the additional implementations listed below.

[0004] The disclosed implementation relates to a hybridization seal used with a microarray that beneficially and advantageously reduces the amount of evaporation that occurs during the hybridization process. The hybridization seal also reduces the possibility that a portion of the microarray dries out due to evaporation of the fluid sample.

[0005] According to the first implementation, the apparatus includes a substrate comprising a plurality of probes and a hybridization encapsulation portion. The hybridization encapsulation portion comprises an evaporation barrier and a layer comprising walls, the walls defining a plurality of sample chambers which form a grid pattern and are for receiving fluid. The layer comprises a first side portion removably bonded to the substrate and a second side portion bonded to the evaporation barrier. The evaporation barrier comprises a barrier section covering the probes and includes one or more slits that allow the barrier section to have a convex or concave profile depending on the amount of fluid in the corresponding sample chamber.

[0006] According to the second implementation, the apparatus includes an evaporation barrier and a layer including walls that support the evaporation barrier and form a grid pattern. The evaporation barrier includes barrier sections, each including an inlet opening and one or more slits.

[0007] According to the third implementation, the apparatus includes a substrate containing a plurality of probes, and a hybridization encapsulation section containing an evaporation barrier and a layer. The evaporation barrier covers the probes and includes barrier sections, each containing a flap and an inlet opening. The layer includes walls, which form a grid pattern and define a plurality of sample chambers for receiving fluid. The layer is removably bonded to the substrate and bonded to the evaporation barrier. The flaps are selectively operable to allow access to the corresponding sample chambers.

[0008] According to the fourth implementation, the method includes removing a liner from a hybridization encapsulation and bonding the hybridization encapsulation to a substrate containing a plurality of probes to form a corresponding sample chamber. The method also includes covering the probes with barrier sections of the evaporation barrier of the hybridization encapsulation. Each barrier section includes an inlet opening and (i) one or more slits or (ii) a flap that can be operated to allow access through the inlet opening.

[0009] According to the fifth embodiment, the method involves dispensing a fluid into a sample chamber through an inlet opening, wherein the sample chamber is located between a substrate containing a probe and a hybridization seal, and the hybridization seal includes an evaporation barrier having a barrier section that covers the probe. The method also includes preventing evaporation of the fluid in the sample chamber by dynamically adjusting the position of the barrier section relative to the substrate based on the amount of fluid in the corresponding sample chamber, or by covering at least a portion of the inlet opening using a flap formed by the barrier section.

[0010] According to the sixth implementation, the method includes removing the liner from the hybridization encapsulation and bonding the hybridization encapsulation to a substrate containing a plurality of probes to form a corresponding sample chamber. The method also includes covering the probes with barrier sections of the evaporation barrier of the hybridization encapsulation. Each barrier section includes an inlet opening and one of the following: (i) one or more slits, (ii) a flap that is operable to allow access through the inlet opening, and (iii) one or more channels.

[0011] According to the seventh implementation, the apparatus includes a substrate and a hybridization encapsulation section. The substrate includes a plurality of probes, and the hybridization encapsulation section includes an evaporation barrier and a layer including walls, the walls defining a plurality of sample chambers which form a grid pattern and are for receiving fluids. The layer includes a first side portion removably bonded to the substrate and a second side portion bonded to the evaporation barrier. The evaporation barrier includes a barrier section that covers the probes and includes one or more channels.

[0012] According to the eighth implementation, the apparatus includes a hybridization encapsulation section, an evaporation barrier, and a layer including a wall, the wall forming a grid pattern and defining a plurality of sample chambers for receiving fluid. The layer includes a first side portion removably bonded to the substrate and a second side portion bonded to the evaporation barrier. The evaporation barrier includes a barrier section for covering a probe and containing one or more channels.

[0013] According to the ninth embodiment, the method involves dispensing a fluid into a sample chamber through an inlet opening, wherein the sample chamber is located between a substrate containing a probe and a hybridization seal, and the hybridization seal includes an evaporation barrier having a barrier section covering the probe. The method also includes flowing a fluid through one or more channels of the corresponding barrier section to allow the fluid to disperse within the corresponding sample chamber.

[0014] According to the tenth implementation, the apparatus includes an evaporation barrier and a layer, the layer including walls forming a grid pattern and including a first side and a second side, the second side being coupled to the evaporation barrier. The evaporation barrier includes a barrier section including one or more channels.

[0015] According to the 11th implementation, the method involves dispensing a fluid into a sample chamber through an inlet opening, wherein the sample chamber is located between a substrate and a hybridization seal, the substrate comprising a probe, and the hybridization seal comprising an evaporation barrier having a barrier section covering the probe. The method also involves flowing a fluid through one or more channels of the corresponding barrier section to disperse the fluid within the corresponding sample chamber.

[0016] According to the twelfth embodiment, the method involves dispensing a fluid into a sample chamber through an inlet opening, wherein the sample chamber is located between a substrate and a hybridization seal, the substrate includes a probe, and the hybridization seal includes an evaporation barrier having a barrier section covering the probe. The method also includes preventing evaporation of the fluid in the sample chamber by dynamically adjusting the position of the barrier section relative to the substrate based on the amount of fluid in the corresponding sample chamber, or by covering at least a portion of the inlet opening using a flap formed by the barrier section.

[0017] Furthermore, according to the first, second, third, fourth, fifth, sixth, seventh, eighth, ninth, tenth, eleventh, and / or twelfth implementations described above, the apparatus and / or method may further include any one or more of the following:

[0018] In one implementation configuration, one or more slits extend from the periphery of the barrier section toward the center of the barrier section.

[0019] In another implementation, one or more slits are at least substantially parallel or at least substantially perpendicular to the wall defining the sample chamber.

[0020] In another implementation, one or more slits are angled relative to the wall defining the sample chamber.

[0021] In another implementation, one or more slits are positioned around at least a portion of each of the barrier sections.

[0022] In another implementation, one or more slits form a barrier sub-section.

[0023] In another implementation, the barrier sub-section has a rectangular shape.

[0024] In another implementation, the barrier sub-section has a triangular shape.

[0025] According to another implementation form, a part of each of the barrier sections is heat-treated.

[0026] According to another implementation form, each of the barrier sections includes an inlet opening.

[0027] According to another implementation form, one or more of the inlet openings are located at different positions of one or more of the barrier sections.

[0028] According to another implementation form, the device includes a first release liner coupled to a first side of the layer and a second release liner covering the evaporation barrier.

[0029] According to another implementation form, the device includes an adhesive on the first side between the first release liner and the layer.

[0030] According to another implementation form, the device further includes a second layer, and an evaporation barrier is coupled between the layer and the second layer.

[0031] According to another implementation form, the evaporation barrier includes a tab extending from between the layer and the second layer.

[0032] According to another implementation form, the flap is arcuate.

[0033] According to another implementation form, the flap is defined by a slit passing through the evaporation barrier.

[0034] According to another implementation form, the slit performs at least one of (i) forming a cross shape and (ii) defining an inlet opening.

[0035] According to another implementation form, it further includes dispensing a fluid into one of the sample chambers and forming a convex profile or a concave profile with a corresponding barrier section based on the amount of fluid in the corresponding sample chamber.

[0036] Another implementation further includes dynamically adjusting the position of the barrier section relative to the substrate based on the amount of fluid in the corresponding sample chamber.

[0037] In another implementation, each barrier section includes one or more slits.

[0038] According to another implementation, the method involves forming a flap using intersecting slits.

[0039] According to another implementation, the method involves forming a flap using an arc-shaped slit.

[0040] In another implementation, the evaporation barrier is substantially rigid.

[0041] In another implementation, the evaporation barrier contains polyethylene terephthalate.

[0042] In another implementation, one or more channels are positioned around at least a portion of each of the barrier sections.

[0043] In another implementation, one or more channels are at least substantially parallel or at least substantially perpendicular to the wall defining the sample chamber.

[0044] In another implementation, the barrier section has a rectangular shape.

[0045] In another implementation, the device further includes a second layer, with an evaporation barrier bonded between the two layers.

[0046] In another implementation, the first layer includes an adhesive multilayer, and the second layer includes a barrier layer.

[0047] According to another implementation, the device further includes a delamination liner bonded to the first side of the layer.

[0048] In another implementation, the apparatus includes a peel liner assembly comprising a peel liner and a peel tab, the peel tab being coupled to the peel liner to allow the peel liner to be removed from the layer.

[0049] In another implementation, the device includes a second peel tab. The peel tab is coupled to the peel liner adjacent to the first end of the peel liner, and the second peel tab is coupled to the peel liner adjacent to the second end of the peel liner.

[0050] In another implementation, the layer has edges that include multiple intersecting surfaces.

[0051] In another implementation, the intersecting surfaces form a sawtooth-like profile.

[0052] According to another implementation, the sawtooth profile allows the layer to be removed more easily from the substrate.

[0053] In another implementation, the edges of the layer define a pair of alignment holes.

[0054] In another implementation, a first sawtooth defines one of the alignment holes at the first edge of the layer, and a second sawtooth defines a second of the alignment holes at the second edge of the layer.

[0055] In another implementation, the layer, the evaporation barrier, and the second layer include alignment holes.

[0056] In another implementation, the alignment holes of the layer, the evaporation barrier, and the second layer are concentric.

[0057] In another implementation, each barrier section includes an entrance opening.

[0058] According to another implementation configuration, the surface of the evaporation barrier facing the substrate includes at least one of having a hydrophilic coating or hydrophilic properties.

[0059] In another implementation, the second layer includes a wall that forms a barrier around the corresponding inlet opening to contain a portion of the fluid protruding from the opening.

[0060] In another implementation, the device includes a frame portion surrounding at least a portion of the evaporation barrier, and further includes a fragile coupling portion between the frame portion and the evaporation barrier.

[0061] In another implementation, each barrier section includes a corner vent.

[0062] In another implementation, the device further includes a second layer, with an evaporation barrier bonded between the two layers.

[0063] In another implementation, the second layer is coupled to the evaporation barrier.

[0064] In another implementation, the second layer and the evaporation barrier are overmolded.

[0065] In another implementation, one or more channels are angled relative to the wall defining the sample chamber.

[0066] In another implementation, each barrier section includes an inlet opening, and one or more channels are connected to the inlet opening.

[0067] In another implementation, the ends of one or more channels are semicircular.

[0068] In another implementation, one or more channels are arranged in a staggered pattern relative to each other.

[0069] In another implementation, a portion of each barrier section is heat-treated.

[0070] In another implementation, heat treatment of a portion of each barrier section allows for deformation that enables the barrier section to have a convex or concave profile depending on the amount of fluid in the corresponding sample chamber.

[0071] In another implementation, multiple probes are coupled to multiple beads, and each of the multiple beads is located inside a sample chamber.

[0072] In another implementation, the second layer includes a wall that forms a barrier around the corresponding entrance opening.

[0073] It should be understood that all combinations of the aforementioned concepts and additional concepts, which are discussed in more detail below (provided that such concepts are not mutually contradictory), are considered to be part of the subject matter of the invention disclosed herein and can be implemented in any combination to achieve the advantages described herein. In particular, all combinations of the claimed subject matter listed at the end of this disclosure are intended to be part of the subject matter of the invention disclosed herein and can be implemented in any combination to achieve the advantages described herein. [Brief explanation of the drawing]

[0074] [Figure 1] A schematic diagram of one implementation of the system as taught in this disclosure is shown. [Figure 2] An isometric view of one implementation configuration of a microarray assembly as taught in this disclosure is shown. [Figure 3] Figure 2 shows a cross-sectional view of the hybridization sealing portion of the microarray assembly. [Figure 4] Figures 2 and 3 show an isometric enlarged view of one mounting configuration of the hybridization encapsulation assembly that can be used with the hybridization encapsulation [Figure 5] Figures 2, 3, and / or 4 show a plan view of one implementation configuration of a barrier section that can be used with the hybridization encapsulation section. [Figure 6]Figures 2, 3, and / or 4 show a plan view of one of the alternative implementations of the barrier section that can be used with the hybridization encapsulation section. [Figure 7] Figures 2, 3, and / or 4 show a plan view of one of the alternative implementations of the barrier section that can be used with the hybridization encapsulation section. [Figure 8] Figures 2, 3, and / or 4 show a plan view of one of the alternative implementations of the barrier section that can be used with the hybridization encapsulation section. [Figure 9] An isometric view of another implementation of the microarray assembly as taught in this disclosure is shown. [Figure 10] Figure 9 shows an isometric view of the hybridization sealing section of the microarray assembly. [Figure 11] Figure 9 shows a cross-sectional view of the hybridization sealing section and the process of dispensing fluid from the sample chamber of the microarray assembly. [Figure 12] Figure 9 shows a cross-sectional view of the hybridization sealing section and the sample chamber of the microarray assembly filled with fluid. [Figure 13A] A plan view is shown of one alternative implementation of a barrier section, including a flap that can be used with the disclosed barrier section. [Figure 13B] A plan view is shown of one alternative implementation of a barrier section, including a flap that can be used with the disclosed barrier section. [Figure 14] This specification shows a flowchart illustrating a method for assembling and using the microarray assemblies disclosed herein. [Figure 15] Another flowchart of a method for assembling the microarray assemblies disclosed herein is shown. [Figure 16] Another flowchart illustrating a method of using the microarray assemblies disclosed herein is shown. [Figure 17] An isometric view of another implementation of the exemplary microarray assembly as taught in this disclosure is shown. [Figure 18]Figure 17 shows an isometric view of the hybridization sealing section of the microarray assembly. [Figure 19] Figures 17 and 18 show an isometric enlarged view of one mounting configuration of the hybridization encapsulation assembly that can be used with the hybridization encapsulation [Figure 20] Figure 19 shows an isometric view of one of the barrier sections of the hybridization sealing assembly. [Figure 21] Figures 17-19 show a plan view of one alternative implementation configuration of the barrier section that can be used to implement the barrier section of the hybridization sealing assembly. [Figure 22] Figures 17-19 show a plan view of one alternative implementation configuration of the barrier section that can be used to implement the barrier section of the hybridization sealing assembly. [Figure 23] Figures 17-19 show a plan view of one alternative implementation configuration of the barrier section that can be used to implement the barrier section of the hybridization sealing assembly. [Figure 24] Figures 17-19 show a plan view of one alternative implementation configuration of the barrier section that can be used to implement the barrier section of the hybridization sealing assembly. [Figure 25] Figures 17-19 show a plan view of one alternative implementation configuration of the barrier section that can be used to implement the barrier section of the hybridization sealing assembly. [Figure 26] Figures 17-19 show a plan view of one alternative implementation configuration of the barrier section that can be used to implement the barrier section of the hybridization sealing assembly. [Figure 27] Figures 17-19 show a plan view of one alternative implementation configuration of the barrier section that can be used to implement the barrier section of the hybridization sealing assembly. [Figure 28] Figures 17-19 show detailed isometric views of alternative mounting configurations for the second layer and evaporation barrier that can be used with the hybridization encapsulation assembly. [Figure 29] Figures 17 and 18 show an isometric enlarged view of one implementation configuration of another hybridization encapsulation assembly that can be used with the hybridization encapsulation [Figure 30] Figures 17 and 18 show an isometric enlarged view of one implementation configuration of another hybridization encapsulation assembly that can be used with the hybridization encapsulation [Figure 31] The process of heat-treating a portion of the barrier section of the evaporation barrier shown in Figure 19 using a tool is illustrated. [Figure 32] Figure 31 shows a schematic cross-sectional view of the barrier section that covers the sample chamber containing the fluid. [Figure 33] Another flowchart illustrating a method of using the microarray assemblies disclosed herein is shown. [Modes for carrying out the invention]

[0075] The following text discloses detailed descriptions of implementations of the method, apparatus, and / or product, but it should be understood that the legal scope of ownership is defined by the claims set out at the end of this patent. Therefore, the following “Modes for Carrying Out the Invention” should be interpreted as illustrative only and do not describe all possible implementations, as it would be impractical, if not impossible, to describe all possible implementations. Numerous alternative implementations may be implemented using either the current art or art developed after the filing date of this patent. Such alternative implementations are expected to still fall within the scope of the claims.

[0076] At least one aspect of the present disclosure is directed to hybridization encapsulation units and associated microarray assemblies. The hybridization encapsulation unit comprises an evaporation barrier and a layer / frame, the layer / frame including walls that form a grid pattern defining a plurality of sample chambers. One side of the layer is removably bonded to a substrate having probes (e.g., a microscope slide, a silicon slide), and the walls form side walls surrounding each probe. The walls positioned around each probe prevent or at least reduce cross-contamination between samples that may be housed in the sample chambers.

[0077] In some implementations, the evaporation barrier may be bonded to the side of a layer facing the substrate to prevent the sample from evaporating during hybridization. The evaporation barrier comprises a barrier section, which has an inlet opening that covers each probe and allows a fluid (e.g., sample) to be loaded into the sample chamber. The inlet opening may be positioned to accommodate the constant spacing of several multichannel pipettes. Thus, the inlet opening may be defined in the center of the barrier section or in another location.

[0078] In some implementations, the barrier section includes one or more slits, which reduce the rigidity of the barrier section, allowing the volume of the sample chamber to be dynamically adjusted, for example, based on the amount of fluid in the sample chamber. Thus, the barrier section may be non-parallel to the substrate and may form a concave or convex profile. In other implementations, the barrier section includes a flap, which is operable to cover the inlet opening and allow the fluid to be loaded into the sample chamber. Regardless of whether the barrier section includes slits or a flap, the disclosed implementations reduce the amount of sample evaporation that may occur during hybridization.

[0079] In other implementations, the height of the sample chamber and the surface tension of the fluid prevent the fluid from dispersing within the sample chamber. To reduce the surface tension of the fluid and / or to allow the fluid to disperse within the sample chamber, in some implementations, the barrier section includes one or more channels.

[0080] Figure 1 shows a schematic diagram of one implementation of System 100 as taught in this disclosure. System 100 can be used to perform analysis on one or more samples of interest. In the shown implementation, System 100 is adapted to receive a substrate 102 containing multiple sites or separate beads 103, which are used to detect the presence of specific target molecules in a sample on the substrate 102. The substrate 102 may be a part of a microarray assembly 200 (see Figure 2) including a hybridization encapsulation 202 (see Figure 2), and / or a microarray assembly (see Figure 17) including a hybridization encapsulation 1701 (see Figure 17), as further disclosed below. System 100 also includes a controller 106 and an imaging system 108. The controller 106 is electrically and / or communicatively coupled to the imaging system 108 and adapted to cause the imaging system 108 to perform various functions disclosed herein.

[0081] The substrate 102 may carry one or more samples of interest (e.g., specimens) and be receptacleable within the substrate receptacle 109. In some implementations, the substrate 102 may contain probe molecules or sites or separate beads 103 at different locations on the substrate 102, each containing a group of probe molecules, allowing for individual access to the probe molecules. For example, one million probes may be provided per bead, and each sample chamber may contain millions of beads (e.g., 10.8 million beads). In other words, each sample chamber may contain thousands, millions, billions, and so on. If beads are provided on the substrate 102, the beads may be coated with DNA oligonucleotides that bind to DNA in the specimen. The sites or separate beads 103 may be referred to as probes 103. In some implementations, the probes are printed on a patch and / or the substrate 102.

[0082] Regardless of how the substrate 102 supports the probe molecules, identifiable labels (e.g., fluorescent markers) are attached to the probe molecules to enable the determination of the presence of a specific target molecule in a sample. In some such implementations, one or more of the identifiable labels have a unique fluorescent label that emits a color when excited. The color (or absence of color) is used to detect the presence of the target molecule in the sample. In the shown implementation, the imaging system 108 is adapted to excite one or more of the identifiable labels (e.g., fluorescent labels) and then acquire image data of this identifiable label. The labels may be excited by incident light and / or laser, and the image data may include one or more colors emitted by each label in response to excitation. The image data (e.g., detection data) may be analyzed by system 100. The imaging system 108 may be a fluorescence spectrophotometer including an objective lens and / or a solid-state imaging device. The solid-state imaging device may include a charge-coupled device (CCD) and / or a complementary metal oxide semiconductor (CMOS).

[0083] Referring to controller 106, in the disclosed implementation, controller 106 includes a user interface 110, a communication interface 112, one or more processors 114, and memory 116, the memory 116 storing instructions executable by one or more processors 114 for performing various functions, including the disclosed implementation. The user interface 110, the communication interface 112, and the memory 116 are electrically and / or communicatively coupled to one or more processors 114.

[0084] In one implementation, the user interface 110 is adapted to receive input from the user and provide the user with information associated with the operation of the system 100 and / or the analysis performed. The user interface 110 may include a touchscreen, display, keyboard, speaker, mouse, trackball, and / or voice recognition system. The touchscreen and / or display may show a graphical user interface (GUI).

[0085] In one implementation, the communication interface 112 is adapted to enable communication between system 100 and a remote system (e.g., a computer) over a network. The network may include the internet, intranet, local area network (LAN), wide area network (WAN), coaxial cable network, wireless network, wired network, satellite network, digital subscriber line (DSL) network, cellular network, Bluetooth connection, or near-field communication (NFC) connection. Some of the communication provided to the remote system may be associated with analysis results, imaging data, etc., generated by system 100 or acquired by other means. Some of the communication provided to system 100 may be associated with fluid analysis operations, patient records, and / or protocols performed by system 100.

[0086] One or more processors 114 and / or system 100 may include one or more processor-based systems or microprocessor-based systems. In some implementations, one or more processors 114 and / or system 100 may include one or more programmable processors, programmable controllers, microprocessors, microcontrollers, graphics processing units (GPUs), digital signal processors (DSPs), reduced-instruction-set computers (RISCs), application-specific integrated circuits (ASICs), field-programmable gate arrays (FPGAs), field-programmable logic devices (FPLDs), logic circuits, and / or other logic-based devices that perform various functions, including those described herein.

[0087] Memory 116 includes semiconductor memory, magnetically readable memory, optical memory, hard disk drive (HDD), optical storage drive, solid-state storage device, solid-state drive (SSD), flash memory, read-only memory (ROM), erasable programmable read-only memory (EPROM), electrically erasable programmable read-only memory (EEPROM), random-access memory (RAM), non-volatile RAM (NVRAM), compact disc (CD), compact disc read-only memory (CD-ROM), digital versatile disk (DVD), Blu-ray disc, and a redundant array of independent disks. This may include one or more of the following storage devices or storage disks: disks, RAID systems, caches, and / or any other storage devices or storage disks on which information is stored for any duration (e.g., permanently, temporarily, over a long period, for buffering, for caching).

[0088] Figure 2 shows an isometric view of one implementation configuration of an exemplary microarray assembly 200 as taught in this disclosure. In the implementation configuration shown, the assembly 200 includes a substrate 102 containing probes 103 and a hybridization encapsulation 202 containing an evaporation barrier 204 and a layer 206. The evaporation barrier 204 may contain a polymer and is shown to be supported by or otherwise bonded to the layer 206. In some implementation configurations, the evaporation barrier 204 includes polyethylene, polyester, polyethylene terephthalate, vinyl, polypropylene, or a combination thereof. The frame 206 may be made of multiple sublayers laminated together, which may be referred to as an adhesive multilayer.

[0089] Layer 206 is removably bonded to the substrate 102 by adhesive 207 and includes walls 208 that form a grid pattern and define a plurality of sample chambers 210. Adhesive 207 may be or may be associated with tapes, double-sided tapes, carriers, glues, acrylic adhesives, silicone adhesives, and / or epoxy that provides a removable watertight seal and is compatible with the reagent used. In the shown configuration, the walls 208 form 24 rectangular sample chambers 210. However, the assembly 200 may include a different number of sample chambers 210, and / or the sample chambers 210 may have different shapes. For example, the assembly 200 or any of the assemblies disclosed herein may include 48 sample chambers 210 or a different number (e.g., 12, 36, or 40) of sample chambers. The walls 208 separating the sample chambers 210 can prevent or reduce cross-contamination of the samples contained in each sample chamber 210.

[0090] The evaporation barrier 204 includes barrier sections 212, each of which covers one of the probe 103 and the sample chamber 210, and can limit the evaporation of the sample during the hybridization step of the assay workflow. In the shown implementation, the barrier section 212 includes one or more slits 214, which allow the barrier section 212 to have a convex or concave profile depending on the amount of fluid in the corresponding sample chamber 210. For example, if a larger amount of fluid is contained in the sample chamber 210, the corresponding barrier section 212 may expand upward and have a convex shape to accommodate the larger fluid volume in the sample chamber 210, and if a smaller amount of fluid is contained in the sample chamber 210, the corresponding barrier section 212 may curve downward and have a concave shape to accommodate the smaller fluid volume in the sample chamber 210. As the barrier section 212 flexes downward, it can facilitate the fluid in the sample chamber 210 to coat the probe 103 and / or distribute the evaporation volume loss over a larger surface area of ​​the probe 103, thereby preventing or delaying the complete evaporation of the fluid, for example, below the inlet opening of the barrier section 212. In other words, the slit 214 increases the flexibility of the barrier section 212 and reduces the adverse effects of evaporation. Additionally or alternatively, the barrier section 212 may undergo heat treatment such as laser ablation or other treatment to increase the flexibility of the barrier section 212 (see, for example, Figures 31 and 32). However, the barrier section 212 may be treated in different ways, which may include applying one or more hydrophobic or hydrophilic treatments to the barrier section 212. Some of these treatments include providing adhesion promoters, coatings, e.g., silicone, silane, phosphate, polyolefin, epoxy, or epoxy, energy treatment, plasma treatment, corona treatment, etching, etc. In other embodiments, the barrier section 212 may be relatively rigid and may be formed of, for example, polyethylene terephthalate (PET), and the slit 214 may be a channel (see, for example, Figures 17 to 31).In some implementations, the channel allows the fluid received in the corresponding sample chamber 210 to disperse within the sample chamber 210, improving the dispersion rate, increasing the dispersion rate, enabling the fluid to disperse within the sample chamber 210, and / or enabling the fluid to disperse within the sample chamber 210.

[0091] In the implementation configuration shown, each barrier section 212 includes one slit 214, the slit 214 being at least substantially parallel to a wall 208 positioned along the longitudinal axis 216 of the hybridization seal 202 and at least substantially perpendicular to a wall 209 positioned along the transverse axis 217 of the hybridization seal 202. As described herein, the phrase “substantially parallel” means being parallel, or not more than about 5° from parallel, and / or taking manufacturing tolerances into consideration, and the phrase “substantially perpendicular” means being perpendicular, or not more than about 5° from perpendicular, and / or taking manufacturing tolerances into consideration. Furthermore, the slit 214 may be formed as a continuous slit intersecting at least one other slit, and / or as a discontinuous slit not intersecting another slit. Although the slit 214 is shown arranged in a particular manner in Figure 2, one or more of the barrier sections 212 may include two or more slits 214, and / or the slits 214 may be arranged differently. For example, the slits 214 may be at least substantially perpendicular to the longitudinal axis 216, and / or one or more slits 214 may extend between the perimeters 218 of the barrier section 212 toward the center 220 of the barrier section 212 (see Figures 4-8). In yet another example, the slits 214 may be angled with respect to the wall 208 defining the sample chamber 210 (Figures 5, 7, 8), and / or the slits 214 may be positioned around each perimeter of the barrier section 212 (see Figures 5, 6, 7). As described herein, the phrase “around the perimeter” means that the slits are located on two or more sides around each perimeter of the barrier section 212. Other arrangements may also be preferred.

[0092] Figure 3 shows an isometric view of the hybridization sealing portion 202 of the microarray assembly 200 of Figure 2. In the shown configuration, the hybridization sealing portion 202 includes an evaporation barrier 204, each having a barrier section 212 that defines a slit 214.

[0093] Figure 4 shows an isometric enlargement of one implementation configuration of a hybridization encapsulation assembly 224 that can be used with the hybridization encapsulation

[0094] In contrast to the layer 206 disclosed in Figures 2 and 3, the hybridization encapsulation assembly 224 in Figure 4 includes layer 206 and a second layer 236, with an evaporation barrier 204 bonded between them. The first layer 206 may be an adhesive, an adhesive multilayer, or a combination of adhesive and polymer film, and the second layer 236 may be a leakage barrier including an adhesive multilayer or a combination of adhesive and polymer film. Furthermore, adhesive may be provided between the first layer 206 and the evaporation barrier 204, between the evaporation barrier 204 and the second layer 236, and / or between the second layer 236 and the second release liner 228. The adhesive may be associated with tape (single-sided, double-sided), glue, etc.

[0095] Referring to the evaporation barrier 204 in Figure 4, in the shown implementation, the evaporation barrier 204 may have a thickness of about 1 / 1000th of an inch (mil) to about 7 mils, and each of the barrier sections 212 includes an inlet opening 240 that allows fluid to be dispensed into the corresponding sample chamber 210. Other thicknesses of the evaporation barrier 204 may also be preferred. The inlet openings 240 may be circular and may be sized to correspond to the outer diameter of the pipette. All inlet openings 240 may be located in the center of the corresponding barrier section 212. Alternatively, the inlet openings 240 may be located, for example, on the barrier section 212, corresponding to the spacing of an eight-tip multichannel pipette.

[0096] The inlet opening 240 exposes the fluid in the sample chamber 210 to the environment, thereby allowing the sample to evaporate at an increased rate below the inlet opening 240. If evaporation occurs, the probe 103 below the inlet opening 240 will dry out, causing a gradient across the probe 103 which may degrade assay performance and / or alter the concentration of DNA in the sample chamber 210. However, advantageously, the slit 214 in the disclosed implementation allows the position of the barrier section 212 to be dynamically adjusted based on the amount of fluid in the sample chamber 210, thereby preventing evaporation.

[0097] The evaporation barrier 204 also includes a tab 242 extending from between layer 206 and a second layer 236, which can be grasped by a person after hybridization in the assay workflow and pulled to separate the hybridization encapsulation 202 from the substrate 102. The evaporation barrier 204 also includes alignment mechanisms 244, 246 of different shapes, which are used to align with the device when assembling the hybridization encapsulation assembly 224 and / or when bonding the hybridization encapsulation assembly 224 to the substrate 102. One of the alignment mechanisms 244 is shown to be circular, and another of the alignment mechanisms 246 is shown to be octagonal. However, the alignment mechanisms 244, 246 may have different shapes and / or may be omitted.

[0098] Figure 5 shows a plan view of one implementation configuration of a barrier section 212 that can be used with the hybridization sealing section 202 of Figures 2, 3, and / or 4. In the implementation configuration shown, the barrier section 212 includes a plurality of slits 214 and an inlet opening 240. The slits 214 in Figure 5 are separate slits 214 in that they do not intersect each other. Thus, the periphery barrier section 248 is connected to the central barrier section 250 via a connecting barrier section 254. The barrier section 212 includes a triangular barrier section 255, which is defined by a periphery slit 256 extending along the periphery of the barrier section 212 and a central slit 258 extending between the barrier section 212 and its center. The central slit 258 is shown extending toward the center of the barrier section 212 at an angle of approximately 45°. However, other angles may also be preferable (e.g., 20°, 27°, 35°), and / or one or more of the slits 214 may have a different angle with respect to the others (see, for example, Figure 8).

[0099] Slits 256 and / or 258 may function as vents for releasing gas (air) from the sample and / or sample chamber 210. If air is present in the sample and / or sample chamber 210, the air may prevent and / or minimize the binding of DNA in the sample to the corresponding probe 103, and in some cases may prevent image data from being obtained from regions of the probe 103 to which DNA did not bind.

[0100] In the shown implementation, the barrier subsection 255 is connected to the periphery barrier section 248 at the corner 260 of the barrier section 212 via the connecting barrier section 254. However, the slits 214 may be arranged differently to allow the periphery barrier section 248 to be connected to the central barrier section 250 at locations other than the corner 260. For example, the slits 214 along each leg around the periphery of the barrier section 212 may include two or more slits 214 (see, for example, Figures 6 and 7), thereby allowing the periphery barrier section 248 to be connected to the central barrier section 250 at locations different from those shown.

[0101] Figure 6 shows a plan view of one implementation configuration of a barrier section 212 that can be used with the hybridization sealing section 202 of Figures 2, 3, and / or 4. In contrast to the barrier section 212 of Figure 5, the slit 214 of the barrier section 212 of Figure 6 is positioned to allow the barrier subsection 255 to have a rectangular shape. The connecting barrier section 254 is located in the center of each leg of the barrier section 212 and extends at least substantially vertically between the periphery barrier section 248 and the central barrier section 250. Other configurations may also be preferred.

[0102] To form a rectangular barrier sub-section 255, each leg of the barrier section 212 includes a pair of peripheral sub-slits 262 and a pair of central sub-slits 264, with a connecting barrier section 254 extending between them. The peripheral sub-slits 262 are coupled to one of the central sub-slits 264, together forming a rectangular barrier section 266 that can form a flat portion of the evaporation barrier 204 that drops down relative to the peripheral barrier section 248. The rectangular barrier section 266 can move outward or inward relative to the substrate 102 to which the hybridization seal 202 is connected, based on the amount of fluid contained in the corresponding sample chamber 210.

[0103] Figure 7 shows a plan view of one implementation configuration of a barrier section 212 that can be used with the hybridization sealing section 202 of Figures 2, 3, and / or 4. In contrast to the barrier sections 212 of Figures 5 and 6, the barrier section 212 includes two perimeter sub-slits 262 on the upper and lower legs 268 and 270 of the barrier section 212, three perimeter sub-slits 262 on the right and left legs 272 and 274 of the barrier section 212, and a central slit 258 connected to the inlet opening 240. In the implementation configuration shown, the barrier section 212 also includes a circular corner vent 267. The corner vent 267 may be used to pass gas from the sample and / or the corresponding sample chamber 210. In other implementation configurations, the corner vent 267 may be in a different location, may have a different shape, or may be omitted.

[0104] Figure 8 shows a plan view of another implementation configuration of one of the barrier sections 212 that can be used with the hybridization sealing section 202 of Figures 2, 3, and / or 4. In contrast to the barrier sections 212 of Figures 5, 6, and 7, the barrier section 212 of Figure 8 does not include a peripheral slit 256 and includes an inlet opening 240 positioned off-center from the center of the barrier section 212 toward the left leg 274, and a central slit 258 extending radially outward from the inlet opening 240 toward the upper leg 268, bottom leg 270, and right leg 272 of the barrier section 212. In the implementation configuration shown, the barrier section 212 also includes a corner vent 267 and a central vent 278 located in the right leg 272 opposite the inlet opening 240.

[0105] Figure 9 shows an isometric view of another implementation of the microarray assembly 300 as taught in this disclosure. The assembly 300 includes a substrate 102 and a hybridization encapsulation section 202 including an evaporation barrier 302 and a layer 206. In contrast to the evaporation barrier 204 disclosed above, the evaporation barrier 302 in Figure 9 does not include a slit 214; instead, each barrier section 212 includes a flap 306 that covers the corresponding inlet opening 240. In the shown implementation, the flap 306 is formed by an arc-shaped slit 308 through the evaporation barrier 302 and a flap connection section 310 that connects the flap 306 to the rest of the barrier section 212. The flap connection section 310 biases the flap 306 toward the covering position. The flap 306 may cover the inlet opening 240 to limit the evaporation of the sample during hybridization in the assay workflow. During operation, the flap 306 is movable between a covered position that covers the inlet opening 240 and an open position that allows access to the corresponding sample chamber 210 and enables the fluid to be dispensed into the sample chamber 210. The flap can be pushed down in different ways, including by using a pipette (see, for example, Figures 11 and 12).

[0106] Figure 10 shows an isometric view of the hybridization sealing section 202 of the microarray assembly 300 shown in Figure 9. In the shown configuration, the hybridization sealing section 202 includes an evaporation barrier 302, each having a barrier section 212 including a flap 306 and an inlet opening 240.

[0107] Figures 11 and 12 show cross-sectional views of the process of dispensing the fluid 314 from the evaporation barrier 302 and sample chamber 210 shown in Figure 9.

[0108] Referring to Figure 11, the pipette 316 is shown to propel the flap 306 into the sample chamber 210 and toward the substrate 102, in the direction generally indicated by arrow 318, thereby enabling access to the sample chamber 210. The pipette 316 dispenses the fluid 314 into the sample chamber 210 through the inlet opening 240. When the pipette 316 is removed, the flap connection portion 310 biases the flap 306 in the direction generally opposite to arrow 318 so as to at least substantially cover the inlet opening 240.

[0109] Figure 12 shows the sample chamber 210 filled with fluid 314, where the flap 306 is at least substantially coplanar with the rest of the evaporation barrier 302 320. As used herein, the term “substantially coplanar” means that the flap 306 is coplanar with the rest of the evaporation barrier 302 320 within about 15°. The sample chamber 210 can accommodate about 15 microliters (μL) and about 20 μL of fluid. However, the sample chamber 210 may be of any other size and / or may accommodate any other amount of fluid depending on the application.

[0110] Figure 13A shows a plan view of one alternative implementation of the barrier section 212, including a flap 306 that can be used with the disclosed barrier section 212. In the example shown, the flap 306 is defined by slits 322 that intersect through the evaporation barrier 204. The slits 322 form four flap sections 324 in a cruciate and triangular shape. It may be apparent that other slit arrangements are preferable for forming the flap 306. For example, a single long slit may be used to form the flap 306, two intersecting slits may be used, and / or three slits that intersect each other at approximately 120°, for example, may be used. In the example shown, the barrier section 212 includes a channel 326, which allows the fluid received in the sample chamber 210 to disperse within the sample chamber 210. For example, channel 326 may allow fluid to flow into channel 326, thereby allowing the fluid to disperse within the sample chamber 210, improving the dispersion rate, increasing the dispersion rate, and / or allowing the fluid to disperse within the sample chamber 210. The fluid may flow along the bottom surface of barrier section 212 and / or along the plane defined by the bottom surface of barrier section 212, and along channel 326. Channel 326 may be referred to as a fluid channel.

[0111] Figure 13B shows a plan view of one alternative implementation of the barrier section 212, which includes flaps 306 that can be used with the disclosed barrier section 212. The barrier section 212 in Figure 13B is similar to the barrier section 212 in Figure 13A. However, in contrast, the barrier section 212 in Figure 13B includes a slit 328, which has a distal end 330 that defines the inlet opening 240 and forms a radial array of flaps 331. The flaps 306 arranged around the inlet opening 240 allow the pipette 316 to engage with one or more of the flaps 306 and push out when dispensing fluid into the sample chamber 210, thereby reducing the possibility of the pipette 316 becoming displaced during the dispensing process.

[0112] Figures 14-16 and 33 show flowcharts of methods using the hybridization encapsulation units 202 and 1701 disclosed herein. The execution order of the blocks may be changed, and / or parts of the described blocks may be modified, deleted, combined, and / or subdivided into multiple blocks.

[0113] The process in Figure 14 begins with the removal of the release liner 226 from the hybridization encapsulation 202 (block 1402). The release liner 226 may be coated with adhesive 207 used to bond the hybridization encapsulation 202 to the substrate 102. The hybridization encapsulation 202 is bonded to the substrate 102, which includes the probe 103, to form the sample chamber 210 and the microarray assembly (block 1404). The sample chamber 210 may be formed by the layer 206 of the hybridization encapsulation 202 and the substrate 102. In the process of bonding the hybridization encapsulation 202 to the substrate 102, the probe 103 is covered with barrier section 212 of the evaporation barrier 204. In some implementations, each of the barrier section 212 includes one or more of the inlet openings 240 and slits 214. In other implementations, each of the barrier sections 212 includes an entrance opening 240 and a flap 306 that is operable to allow access through the entrance opening 240.

[0114] In block 1408, fluid is dispensed into one of the sample chambers 210, and the corresponding barrier section 212 forms a convex or concave profile based on the amount of fluid in the corresponding sample chamber 210 (block 1408). The position of the barrier section 212 is dynamically adjusted relative to the substrate 102 based on the amount of fluid in the corresponding sample chamber 210. The microarray assembly then undergoes a hybridization process, after which the hybridization encapsulation section 202 is removed from the substrate 102, and the substrate 102 is imaged using system 100 to identify target molecules of interest in the sample on the substrate 102.

[0115] The process in Figure 15 begins with the removal of the peel liner 226 from the hybridization encapsulation 202 (block 1502). The hybridization encapsulation 202 is bonded to a substrate 102 containing the probe 103 to form the sample chamber 210 and the microarray assembly (block 1504). The sample chamber 210 may be formed by the layer 206 of the hybridization encapsulation 202 and the substrate 102. In the process of bonding the hybridization encapsulation 202 to the substrate 102, the probe 103 is covered with a barrier section 212 of the evaporation barrier 204. In some implementations, each of the barrier section 212 includes one or more of the inlet openings 240 and slits 214. In other implementations, each of the barrier section 212 includes an inlet opening 240 and a flap 306 that is operable to allow access through the inlet opening 240. In other implementations, each of the barrier sections 212 includes an inlet opening 240 and one or more channels (see, for example, Figures 13A, 13B, and 17-29).

[0116] The method in Figure 16 begins with the dispensing of a fluid into a sample chamber 210 through an inlet opening 240 (block 1602). The sample chamber 210 is located between a substrate 102 and a hybridization seal 202, the substrate 102 containing a probe 103, and the hybridization seal 202 containing an evaporation barrier 204 having a barrier section 212 covering the probe 103. Evaporation of the fluid in the sample chamber 210 is prevented by dynamically adjusting the position of the barrier section 212 relative to the substrate 102 based on the amount of fluid in the corresponding sample chamber 210, or by covering the inlet opening 240 using a flap 306 formed by the barrier section 212. Each of the barrier sections 212 may contain one or more slits 214, which may be intersecting slits 322 or arc-shaped slits 308.

[0117] Figure 17 shows an isometric view of another implementation of the microarray assembly 1700 as taught in this disclosure. Assembly 1700 is similar to assembly 200 in Figure 2. However, in contrast to the evaporation barrier 204 which includes slits 214, the microarray assembly 1700 includes a hybridization encapsulation section 1701 which has an evaporation barrier 1702 which includes barrier sections 1704, each containing more channels 1706. The barrier sections 1704 cover one of the probes 103 and the sample chamber 210, which can limit the evaporation of the sample during the hybridization step of the assay workflow.

[0118] In the disclosed implementation, the height 1707 between the substrate 102 and the evaporation barrier 1702 may be such that the surface tension of the fluid deposited in the sample chamber 210 prevents the fluid from dispersing within the sample chamber 210. The height 1707 may be about 240 micrometers (μm) to about 500 micrometers or another distance. Using the disclosed implementation, the channel 1706 is positioned and / or configured to reduce the surface tension of the fluid, to allow the dispersion of the fluid within the corresponding sample chamber 210, to improve the fluid flow within the corresponding sample chamber 210, and / or to improve the distribution of the fluid within the corresponding sample chamber 210. To further facilitate the dispersion of the fluid within the sample chamber 210, the surface of the evaporation barrier 1702 facing the substrate 102 may include a hydrophilic coating, and / or the evaporation barrier 1702 may have hydrophilic properties. To further facilitate the fluid entering, for example, the corresponding inlet opening of the evaporation barrier 1702 and into the sample chamber 210, the surface of the evaporation barrier 1702 facing away from the substrate 102 may have lower hydrophilic properties and / or hydrophobic properties, and / or may include a hydrophobic layer (e.g., a thin transparent plastic layer).

[0119] In the shown configuration, the channel 1706 is shown to be at least substantially parallel to the wall 209 positioned along the longitudinal axis 216 of the hybridization seal 1701 and at least substantially perpendicular to the wall 209 positioned along the transverse axis 217 of the hybridization seal 202. However, the channel 1706 may be positioned differently. In the shown configuration, the wall 208 forms 24 rectangular sample chambers 210, and the evaporation barrier 1704 may include polyethylene, polyester, polyethylene terephthalate, vinyl, polypropylene, or a combination thereof. However, the assembly 1700 may include a different number of sample chambers 210, the sample chambers 210 may have different shapes, and / or different materials may be used.

[0120] Although the channels 1706 are shown arranged in a particular manner in Figure 17, one or more of the barrier sections 1704 may contain two or more channels 1706, and / or the channels may be arranged differently. For example, the channels 1706 may be at least substantially perpendicular to the longitudinal axis 216, and / or one or more channels 1706 may extend from the perimeter 218 of the barrier section 1704 toward the center 220 of the barrier section 212 (see Figures 19-27). In yet another example, the channels 1706 may be angled with respect to the wall 208 defining the sample chamber 210 (see Figures 21-24 and 26 and 27), and / or the channels 1706 may be positioned around each of the perimeters of the barrier section 212 (see Figures 21-24 and 26 and 27).

[0121] Additionally or alternatively, the barrier section 1704 may undergo heat treatment, such as laser ablation, or other treatment to increase the flexibility of the barrier section 1704. In some implementations, heat treatment of the barrier section 1704 allows the barrier section 170 to deform depending on the amount of fluid in the corresponding sample chamber 210, thereby having a convex or concave profile, thereby preventing or delaying the complete evaporation of the fluid, for example, below the inlet opening of the barrier section 1704. In some such implementations, the barrier section 1704 may be selectively heat-treated such that each central portion 1708 of the barrier section 1704 has increased flexibility, and each peripheral portion 1710 of the barrier section 1704 has relatively higher rigidity than the central portion 1708.

[0122] Figure 18 shows an isometric view of the hybridization encapsulation section 1701 of the microarray assembly 1700 of Figure 17. In the shown configuration, the hybridization encapsulation section 1701 includes an evaporation barrier 1702, each having a barrier section 1704 containing a channel 1706.

[0123] Figure 19 shows an isometric enlargement of one implementation configuration of a hybridization encapsulation assembly 1750 that can be used with the hybridization encapsulation The adhesive 207 may be adapted to form a watertight seal with the substrate 102 when the hybridization seal 1701 is bonded to the substrate 102, while also being adapted to allow the hybridization seal 1701 to be separated from the substrate 102. The adhesive 207 may be or be associated with a tape, double-sided tape, glue, etc.

[0124] The peel tab 1756 is bonded to the first release liner 1752 at its end 1758, allowing the first release liner 1752 to be removed from layer 206. The release liner 1752 and the peel tab 1756 form a release liner assembly 1757. The peel tab 1756 may be bonded to the first release liner 1752 using permanent adhesive or another type of fastener. Alternatively, the peel tab 1756 may be integral with the first release liner 1752. Although the first release liner 1752 is shown with a single peel tab 1756 bonded to it, it may include two or more peel tabs (see, for example, Figure 30), or the peel tab 1756 may be omitted. In a configuration including two peel tabs 1756, one of the peel tabs 1756 may be coupled to an end 1758, and the other peel tab 1756 may be coupled to the other end 1764 of the first peel liner 1752. If the peel tabs 1756 are omitted, the first peel liner 1752 may be sized larger than the layer 206 to allow the first peel liner 1752 to be easily grasped and removed from the layer 206 by an operator.

[0125] Similar to the hybridization encapsulation assembly 224 in Figure 4, the hybridization encapsulation assembly 1750 in Figure 19 includes a layer 206 and a second layer 1762 to which an evaporation barrier 1702 is bonded. Layer 206 may be an adhesive, an adhesive multilayer, or a combination of an adhesive and a polymer film, and the second layer 1762 may be a leakage barrier including an adhesive multilayer or a combination of an adhesive and a polymer film. Furthermore, adhesive may be provided between layer 206 and the evaporation barrier 1702, between the evaporation barrier 1702 and the second layer 1762, and / or between the second layer 1762 and the second release liner 1754.

[0126] In the shown configuration, the end 1767 of layer 206 includes a plurality of intersecting surfaces 1766 that form a sawtooth profile 1768. Advantageously, the sawtooth profile 1768 allows force to be concentrated at the end 1770 of the sawtooth profile 1768 when the hybridization seal 1701 is being removed from the substrate 102, thereby potentially preventing the sample and / or fluid from inadvertently splashing from the substrate 102 during the removal process. In other words, the sawtooth profile 1768 allows layer 206 to be removed more easily from the substrate 102. In the shown configuration, the central teeth 1772 of the sawtooth profile 1768 may be similar and / or identical, and the outer teeth 1774 may be similar, identical, or mirror images of each other.

[0127] Referring further to layer 206, the end 1767 of layer 206 defines a pair of alignment holes 1778, one of which is defined by one of the outer teeth 1774, and the other of which is defined by the other of the outer teeth 1774. The evaporation barrier 1702 and the second layer 1762 also include alignment holes 1780, 1782. Thus, layer 206, evaporation barrier 1702, and the second layer 1762 include alignment holes 1178, 1780, 1782 which can be used to align layer 206, evaporation barrier 1702, and the second layer 1762 with respect to each other, for example, during the assembly and / or manufacturing process. In other words, the alignment holes 1178, 1780, 1782 can facilitate the alignment between layers.

[0128] When layer 206, evaporation barrier 1702, and second layer 1762 are joined, the alignment holes 1178, 1780, and 1782 may be concentric with each other, allowing for easy inspection of the alignment of layer 206, evaporation barrier 1702, and second layer 1762. The alignment holes 1178, 1780, and 1782 are shown to be circular. However, the alignment holes 1178, 1780, and 1782 may have other shapes and / or be different from each other. Alternatively, other types of alignment mechanisms may be included. To allow for easy visual inspection of whether the alignment mechanism is aligned, for example, one of the alignment holes 1178, 1780, and 1782 may be square and / or circular, and another of the alignment holes 1178, 1780, and 1782 may be cross-shaped and / or crosshair-shaped.

[0129] In the provided implementation, referring to the evaporation barrier 1702, each of the barrier sections 1704 of the evaporation barrier 1702 includes an inlet opening 240 that allows fluid to be dispensed into the corresponding sample chamber 210. The inlet opening 240 may be circular, rectangular, and / or squircle-shaped (intermediate between square and circular) and may be sized to correspond to the outer diameter of the pipette. In some implementations, the inlet opening 240 may be positioned to accommodate a certain interval between several multichannel pipettes. Thus, the inlet opening 240 may be defined in the center or elsewhere within the barrier section 1704.

[0130] The evaporation barrier 204 also includes a tab 242 extending from between the layer and the second layer 1762, which can be grasped by a person after hybridization in the assay workflow and pulled to separate the hybridization seal 1701 from the substrate 102. The evaporation barrier 1702 also includes a frame portion 1784 surrounding at least a portion of the evaporation barrier 1702, and includes a fragile joint 1786 between the frame portion 1784 and the evaporation barrier 1702. The fragile joint 1786 may be a tab and / or perforation that allows the frame portion 1784 to be removed from around the evaporation barrier 1702 when broken. The frame portion 1784 may be removed after the hybridization seal 1701 has been bonded to the substrate 102.

[0131] Referring to the second layer 1762, the second layer 1762 includes a wall 1788 that forms a barrier 1790 around the corresponding inlet opening 240. The barrier 1790 may serve to contain any portion of the fluid (e.g., a convex meniscus) that may protrude from the inlet opening 240 after the fluid has been dispensed into the inlet opening 240.

[0132] Figure 20 shows a plan view of one of the barrier sections 1704 of the hybridization encapsulation section 1701 in Figure 19. In the shown configuration, the barrier section 1704 includes an inlet opening 240 and a plurality of channels 1706. The inlet opening 240 is shown as a squircle shape. However, the inlet opening 240 may have any shape (e.g., circular, rectangular).

[0133] The channels 1706 in Figure 20 are separate channels 1706 in that they do not intersect with each other. Specifically, barrier section 1704 includes three channels 1706 in the upper 1792 and bottom 1794 of barrier section 1704, and barrier section 1704 includes four channels 1706 in the left side 1796 and right side 1798 of barrier section 1704. Barrier section 1704 also includes a corner vent 1800 which may allow gas (air) to be released from the sample and / or sample chamber 210 and / or facilitate the dispersion of the sample within the sample chamber 210. In the shown configuration, the spacing between channels 1706, the spacing between channels 1706 and the wall 1788, and the spacing between channels 1706 and the corner vent 1800 facilitate the dispersion of the sample within the sample chamber 210. In some implementations, to facilitate the dispersion of the sample within the sample chamber 210, the channels 1706 are spaced at a first distance 1802, and the channels 1706 and the corner vents 1800 are spaced at a second distance 1804 that is different from (e.g., greater than) the first distance 1802. Furthermore, to facilitate the dispersion of the sample within the sample chamber 210, the corner vents 1800 may be offset with respect to the axis 1806 of the channels 1706. Each corner 1802 of the barrier section 1704 includes a corresponding corner vent 1800, although one or more of the corners 1802 may include a corner vent 1800 or may omit the corner vent 1800.

[0134] Figure 21 shows a plan view of one of the barrier sections 1850 that can be used to implement the barrier section 1704 of the hybridization sealing section 1701 shown in Figures 17-19, in a different implementation configuration. In the implementation configuration shown, the barrier section 1850 includes a plurality of channels 1706 which are central channels 1852, these central channels 1852 are connected to or otherwise coupled with the inlet opening 240 and spaced apart from the perimeter 218 of the barrier section 1850.

[0135] Some of the central channels 1852 are at least substantially parallel or at least substantially perpendicular to the sides 1792, 1794, 1796, and 1798 of the barrier section 1850, and some of the central channels 1852 are angled with respect to the sides 1792, 1794, 1796, and 1798 of the barrier section 1850. The angled channels are shown to have ends 1854 with an enlarged opening 1856 with respect to the width of the rest of the corresponding channel 1852. As shown, the enlarged opening 1856 is semicircular, and therefore some ends 1854 of the central channels 1852 are semicircular. However, the enlarged opening 1856 may have a different shape.

[0136] Figure 22 shows a plan view of one of the barrier sections 1900 that can be used to implement the barrier section 1704 of the hybridization encapsulation section 1701 in Figures 17-19, but with the central channel 1852, which is at least substantially parallel or at least substantially perpendicular to the sides 1792, 1794, 1796, and 1798 of the barrier section 1850, omitted.

[0137] Figure 23 shows a plan view of one of the barrier sections 1950 that can be used to implement the barrier section 1704 of the hybridization encapsulation section 1701 in Figures 17-19, in a different implementation configuration. The barrier section 1950 in Figure 23 is similar to the barrier section 1900 in Figure 22. However, in contrast, the inlet opening 240 of the barrier section 1950 in Figure 23 has sides of different lengths, such that the inlet opening 240 is elliptical.

[0138] Figure 24 shows a plan view of one of the barrier sections 2000 that can be used to implement the barrier section 1704 of the hybridization encapsulation section 1701 in Figures 17-19, but in contrast, the central channel 1852 of the barrier section 1950 does not include the enlarged opening 1856.

[0139] Figure 25 shows a plan view of another implementation configuration of one of the barrier section 2500s that can be used to implement the barrier section 1704 of the hybridization encapsulation section 1701 in Figures 17-19. The barrier section 2500 in Figure 25 is similar to the barrier section 1850 in Figure 21. However, in contrast, the barrier section 2500 in Figure 25 includes a corner vent 1800 and staggered channels 2502 relative to each other. The staggered channels 2502 are at least substantially parallel or at least substantially perpendicular to the sides 1792, 1794, 1796, and 1798 of the barrier section 1850. In the implementation configuration shown, three of the channels 2502 are positioned between the inlet opening 240 and the top 1792 of the barrier section 2500, and three of the channels 2502 are positioned between the inlet opening 240 and the bottom 1794 of the barrier section 2500. However, any number of channels 2502 may be included.

[0140] Figure 26 shows a plan view of one of the barrier sections 2550, an alternative implementation configuration, that can be used to implement the barrier section 1704 of the hybridization encapsulation section 1701 in Figures 17-19. The barrier section 2550 in Figure 26 is similar to the barrier section 1850 in Figure 21. However, in contrast, the channels 1706 angled relative to the sides 1792, 1794, 1796, and 1798 of the barrier section 1850 are staggered channels 2502 that are not connected to the inlet opening 240.

[0141] Figure 27 shows a plan view of one of the barrier sections 2600 that can be used to implement the barrier section 1704 of the hybridization sealing section 1701 in Figures 17-19, but with the exception of the barrier section 1850 in Figure 21.

[0142] Figure 28 shows a detailed isometric view of an alternative mounting configuration of the second layer 1762 and evaporation barrier 1702 that can be used with the hybridization encapsulation 1701 of Figures 17-19. In the mounting configuration shown, the second layer 1762 includes a wall 1788, which, together with the evaporation barrier 1702, forms a barrier 1790 around the inlet opening 240. The evaporation barrier 1702 includes a channel 2650 that extends from the wall 1788 and facilitates manufacturability. Although the channel 2650 is shown to be included, it may be omitted in other mounting configurations.

[0143] Figure 29 shows an isometric enlargement of one implementation configuration of another hybridization encapsulation assembly 2700 that can be used with the hybridization encapsulation assembly 1701 of Figures 17 and 18. The hybridization encapsulation assembly 2700 of Figure 29 is similar to the hybridization encapsulation assembly 1750 of Figure 19. However, in contrast, the hybridization encapsulation assembly 2700 of Figure 29 has more barrier sections 1704, and the frame 206 has walls 208 that form a corresponding number of sample chambers 210. In the implementation configuration shown, 48 barrier sections 1704 are provided. However, any number of barrier sections may be included. Furthermore, in contrast to the hybridization encapsulation assembly 1750 of Figure 19, the layer 206 and the second layer 1762 include tabs 242, and the frame portion 1784 surrounding the evaporation barrier 1702 includes alignment holes 1780, in contrast to the evaporation barrier 1702 itself including alignment holes. Furthermore, while layer 206 in Figure 29 does not include intersecting surfaces 1766 that form the sawtooth profile 1768, layer 206 in Figure 29 may alternatively include intersecting surfaces 1766 that form the sawtooth profile 1768.

[0144] Figure 30 shows an isometric enlargement of one implementation configuration of another hybridization encapsulation assembly 2800 that can be used with the hybridization encapsulation encapsulation 1701 of Figures 17 and 18. The hybridization encapsulation assembly 2800 of Figure 30 is similar to the hybridization encapsulation assembly 2700 of Figure 29. However, in contrast, the hybridization encapsulation assembly 2800 of Figure 30 includes a second layer 1762 bonded to the evaporation barrier 1702 by, for example, an overmolding process. The overmolding process may include a silicone injection overmolding process. However, the evaporation barrier 1702 and the second layer 1762 can be bonded in any preferred way. By bonding the second layer 1762 and the evaporation barrier 1702 by overmolding, alignment problems between the second layer 1762 and the evaporation barrier 1702 can be eliminated or reduced. In some implementations, the second layer 1762 is made of rubber such as silicone rubber, and the evaporation barrier 1702 is made of a relatively soft, overmold plastic. However, the second layer 1762 and / or the evaporation barrier 1702 can be made of any suitable material.

[0145] In addition, in contrast to the hybridization encapsulation assembly 2700 in Figure 29, the layer 206 in Figure 30 includes two peel tabs 1756 arranged to be bonded to the layer 206 at opposing ends 2802, 2803 of the layer 206. Alternatively, a single peel tab 1756 may be included, or the peel tab 1756 may be omitted.

[0146] Figure 31 shows the process of heat-treating portion 2804 of the barrier section 1704 of the evaporation barrier in Figure 19 using tool 2852. Tool 2852 may be a heated mechanical stamp. Advantageously, heat-treating portion 2804 allows the barrier section 1704 to have a convex or concave profile depending on the amount of fluid in the corresponding sample chamber 210. In other words, locally heat-treating the barrier section 1702 allows the barrier section 1702 to maintain contact with the fluid in the sample chamber 210, thereby minimizing contact between the fluid and air and reducing the evaporation rate. In the shown implementation, the barrier section 1702 may be made of PET, has a relatively rigid perimeter 2854, and includes a channel 1706. Alternatively, the channel 1706 may be omitted.

[0147] Figure 32 shows a schematic cross-sectional view of the barrier section 1704 of Figure 31, covering the sample chamber 210 containing the fluid 2856. In the implementation configuration based on the locally heat-treated / stressed portion 2804 of the barrier section 1704 shown, portion 2804 forms a convex / volcanic profile. Conversely, as the fluid 2856 evaporates, portion 2804 can move in the direction generally indicated by arrow 2858, thereby allowing contact between the barrier section 1704 and the fluid 2856 to be maintained.

[0148] The method in Figure 33 begins by dispensing a fluid dispensed through an inlet opening 240 into a sample chamber 210 (block 3302). The sample chamber 210 is located between a substrate 102 and a hybridization seal 1701, the substrate 102 containing a probe 103, and the hybridization seal 1701 containing an evaporation barrier 1702 having a barrier section 1704 covering the probe 103. Fluid is flowed through one or more channels 1706 of the corresponding barrier section 1704 to disperse the fluid within the corresponding sample chamber 210 (block 3304). In some implementations, flowing fluid through one or more channels 1706 increases and / or maximizes the amount of fluid covering the substrate 102. In some implementations, the channels 1706 are angled with respect to a wall 1788 defining the sample chamber 210. In some implementations, the channels 1706 are positioned around at least a portion of each perimeter 218 of the barrier section 1704. In other implementations, one or more channels 1706 are at least substantially parallel or at least substantially perpendicular to the wall 1788 that defines the sample chamber 210.

[0149] The foregoing description is provided to enable those skilled in the art to implement the various configurations described herein. While the subject art has been described in particular with reference to various figures and configurations, it should be understood that these are for illustrative purposes only and should not be construed as limiting the scope of the subject art.

[0150] When used herein, an element or process described in the singular and followed by the word "a" or "an" should be understood not to exclude multiple such elements or processes unless such exclusion is explicitly stated. Furthermore, a reference to "one implementation" is not intended to be interpreted as excluding the existence of additional implementations that also incorporate the listed features. Moreover, unless explicitly stated otherwise, an implementation that "comprises," "includes," or "having" one or more elements having a particular characteristic may include additional elements, whether or not they possess that characteristic. Furthermore, terms such as "comprising," "including," and "having" are used interchangeably herein.

[0151] The terms “substantially,” “approximately,” and “about” as used throughout this specification are used to describe and account for small variations resulting from processing variability, etc. For example, they may refer to ±5% or less, e.g., ±2% or less, e.g., ±1% or less, e.g., ±0.5% or less, e.g., ±0.2% or less, e.g., ±0.1% or less, e.g., ±0.05% or less.

[0152] Many other methods may exist for carrying out the subject art. Various functions and elements described herein may be divided in ways different from those shown without departing from the scope of the subject art. Various modifications to these implementations may be readily apparent to those skilled in the art, and the general principles defined herein may be applicable to other implementations. Thus, many changes and modifications may be made to the subject art by those skilled in the art without departing from the scope of the subject art. For example, a different number of a given module or unit may be used, different or more types of a given module or unit may be used, a given module or unit may be added, or a given module or unit may be omitted.

[0153] Underlined and / or italicized headings and subheadings are used solely for convenience and do not limit the subject art, nor are they referenced in connection with the interpretation of the description of the subject art. All structural and functional equivalents of elements of various implementations described throughout this disclosure, which are known to those skilled in the art or will become known thereafter, are expressly incorporated herein by reference and are intended to be included in the subject art. Furthermore, nothing disclosed herein is intended to be for publication only, whether such disclosure is expressly stated in the above description or not.

[0154] It should be understood that all combinations of the aforementioned concepts and further concepts, which will be discussed in more detail below (on the premise that such concepts are not contradictory), are considered to be part of the subject matter disclosed herein. Specifically, all combinations of the claimed subject matter appearing at the end of this disclosure are considered to be part of the subject matter disclosed herein.

Claims

1. It is a device, A substrate containing multiple probes, It comprises a hybridization sealing section, and the hybridization sealing section is Evaporation barrier and A layer including walls, wherein the walls define a plurality of sample chambers that form a grid pattern and are for receiving fluids, It has a second layer, The layer includes a first side portion removably bonded to the substrate and a second side portion bonded to the evaporation barrier, The evaporation barrier is bonded between the first layer and the second layer. The apparatus wherein the evaporation barrier covers the probe and includes a barrier section containing one or more channels.

2. The apparatus according to claim 1, wherein the evaporation barrier is rigid.

3. The apparatus according to claim 1 or 2, wherein one or more channels are positioned around at least a portion of each of the barrier sections.

4. The apparatus according to any one of claims 1 to 3, wherein one or more channels are parallel or perpendicular to the wall defining the sample chamber.

5. The apparatus according to any one of claims 1 to 4, wherein the aforementioned layer includes an adhesive multilayer, and the second layer includes a barrier layer.

6. The apparatus according to any one of claims 1 to 5, wherein the surface of the evaporation barrier facing the substrate is a hydrophilic coating or has hydrophilic properties, the apparatus according to any one of these.

7. The apparatus according to any one of claims 1 to 6, wherein the layer has an end having a plurality of intersecting surfaces, and the intersecting surfaces form a sawtooth profile.

8. The apparatus according to any one of claims 1 to 7, wherein the layer, the evaporation barrier, and the second layer are provided with alignment holes.

9. The apparatus according to claim 8, wherein the alignment holes of the layer, the evaporation barrier, and the second layer are concentric.

10. It is a device, Evaporation barrier and Layers, It has a second layer, The layer includes walls forming a grid pattern and includes a first side and a second side, the second side being coupled to the evaporation barrier. The evaporation barrier is bonded between the first layer and the second layer. The apparatus wherein the evaporation barrier includes a barrier section containing one or more channels.

11. The apparatus according to claim 10, wherein each of the barrier sections includes an inlet opening, and the second layer includes a wall that forms a barrier around the corresponding inlet opening.

12. The apparatus according to claim 10 or 11, wherein each of the barrier sections includes a corner vent.

13. The apparatus according to any one of claims 10 to 12, wherein the second layer and the evaporation barrier are overmolded.

14. The apparatus according to any one of claims 10 to 13, wherein each of the barrier sections includes an inlet opening, and one or more channels are coupled to the inlet opening.

15. The apparatus according to claim 14, wherein the ends of one or more channels are semicircular.

16. The apparatus according to any one of claims 10 to 14, wherein one or more of the channels are arranged in a staggered pattern relative to each other.

17. The apparatus according to any one of claims 10 to 16, wherein a portion of each of the barrier sections is heat-treated, and the heat treatment of the portion of each of the barrier sections allows for deformation that enables the barrier section to have a convex or concave profile depending on the amount of fluid in the corresponding sample chamber.

18. The layer further comprises a release liner bonded to the first side of the aforementioned layer, The apparatus according to any one of claims 10 to 17, further comprising a peel liner assembly comprising the peel liner and a peel tab, wherein the peel tab is coupled to the peel liner to enable the peel liner to be removed from the layer.

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