CN217033310U - Supporting device - Google Patents

Abstract

A support device includes a substrate holder including a cover configured to receive a pad, a base configured to receive a sample substrate, and first and second locking tabs. The first and second locking tabs include a movable tab coupled to the opposing first sidewall of one of the cover and the base and a non-movable tab coupled to the opposing first sidewall of the other of the cover and the base. The length of the first locking tab is greater than the length of the second locking tab. The first locking tab extends along a length of the liner. The movable tab is configured to engage with the non-movable tab to releasably secure the cover to the base.

Description

Supporting device

Technical Field

The present disclosure relates to an analysis device, in particular a support device, for a biological sample.

Background

Cells within the tissue being tested may differ in cell morphology and/or function due to different analyte levels (e.g., gene and/or protein expression) within different cells. The particular location of a cell within a tissue (e.g., the location of a cell relative to neighboring cells or the location of a cell relative to the tissue microenvironment) may affect, for example, the morphology, differentiation, fate, viability, proliferation, behavior, and signaling of cells, as well as cross-talk with other cells in the tissue.

Spatial heterogeneity has previously been studied using techniques that provide data for only a small amount of analyte in a whole or partial tissue, or provide data for a large amount of analyte in a single cell, but fail to provide information about the location of a single cell in a parental biological sample (e.g., a tissue sample).

Furthermore, the resolution and sensitivity of the imaging system for spatial analyte data is inherently variable. This is due in large part to the variability of manufacturers of imaging system components, in addition to the differences between the arrangement of the imaging devices, the various types of imaging devices, and the image acquisition software. The image quality is also affected by changes in the image acquisition performed by the user. This problem becomes more pronounced when attempting to image a sample of unknown fluorescence intensity or when imaging a sample by a user of different experience.

Furthermore, in a laboratory environment, various processing protocols (protocols) are used to prepare samples for analysis. These protocols can be carried out in test tubes, on slides, or more generally on samples supported by a matrix. Certain protocols are performed at stable, controlled temperatures to ensure fidelity of the sample and protocol reagents. Other protocols involve temperature cycling and other steps in which the temperature of the sample is adjusted in a controlled manner. To heat the sample and its supporting substrate during the protocol, a thermal cycler, hot plate, or other heating device may be used. As an example, a thermal cycler can be used for nucleic acid amplification and sequence analysis by transcription and reverse transcription as part of a polymerase chain reaction protocol. The samples may also be subjected to controlled heating in thermocyclers and other heating devices, for example to facilitate temperature sensitive reactions for restriction enzyme digestion and rapid diagnosis.

In addition, biological samples may be placed on a solid support for analysis to identify or characterize analytes within the sample, such as DNA, RNA, or other genetic material. The printed guide may help improve the placement of the sample on the solid support.

Current methods and devices for supporting biological samples (e.g., during spatial analysis and/or heating) often lack portability, do not provide access to specific sample areas or wells, and/or do not provide hermetic seals for preventing cross-contamination between sample areas or wells.

SUMMERY OF THE UTILITY MODEL

Embodiments disclosed below include a support device for a substrate including a sample region and a method of incubating a sample disposed on the sample region of the substrate. In some embodiments, a distinguishing advantage of the support apparatus and method of the present disclosure is that it is configured as a one-piece device that provides ease of use for the user. For example, the one-piece design facilitates setup and reduces the time spent by the user in assembling the device and the base. In some embodiments, a user may easily insert a substrate into the described device without fastening multiple components. In some embodiments, the user does not use any tools to assist in inserting the substrate into the device or to assist in removing the substrate from the device. For example, a user may not need to use any type of fastener to assemble the support device or secure the base to the support device, thereby providing a more efficient way to support the biological sample. In other examples, the devices disclosed herein may alternatively use one or more fasteners to assemble and/or secure the base to the support device.

In some embodiments, another distinct advantage of the support apparatus and methods of the present disclosure is its design that provides access to a particular sample area in a substrate through the alignment of a plurality of holes in a substrate holder and a plurality of openings in a gasket. In some embodiments, an additional advantage of the support apparatus and method of the present disclosure is the uniform pressure applied to the substrate via the gasket and/or rib or ribs when the support apparatus is in the closed position. Further, a hermetic seal and/or a gas seal is formed between the gasket and the substrate when the substrate holder is in the closed position. Such hermetic and/or gas-tight seals may prevent fluid transfer between the plurality of openings of the gasket. The hermetic seal and/or gas seal may further prevent cross-contamination of the biological sample via, for example, leakage of a solution or fluid from the first sample region to the second sample region. In still other embodiments, a further advantage of the support apparatus and methods described in this disclosure is that shear forces on the substrate, e.g., on the section of the substrate proximate to the one or more hinges, are minimized. The minimization of shear forces may prevent damage (e.g., breaking or cracking) to the substrate.

The apparatus provided herein can provide consistent and uniform heating of a substrate surface. Even heating may be critical to ensure that the preparative reactions performed on the substrate-supported samples occur according to established protocols and achieve the desired results.

Further, if the substrate is heated without the cover, heating the substrate (e.g., glass slide) to a temperature above room temperature may result in the formation of condensate on the upper surface of the enclosed substrate wells. The condensate may alter the composition of the reaction mixture in the base wellbore, inhibit the production reaction, and/or produce unpredictable results. The devices described in this disclosure may be used to reduce or prevent the formation of condensate in the basewellbore by the gas-tight seal and/or gas seal created by the liner.

Certain types of thermal cyclers and heating devices are specifically constructed for a particular type of substrate (e.g., a multi-well substrate). Loading other types of substrates (e.g., standard microscope slides) into such devices may result in non-uniform heating of the substrate. The apparatus described in the present disclosure may be used to support a substrate within a heating apparatus that is not designed for such a substrate, thereby ensuring adequate and uniform heat transfer to the substrate. In particular, these devices can be used to accommodate thermal cyclers designed to accept multi-well substrates, such that other types of substrates can be effectively heated within the thermal cycler as part of a sample preparation protocol.

In some embodiments, the devices of the present disclosure allow the surface of the substrate to directly contact the surface of a heating device (e.g., a thermal cycler), thereby allowing uniform heating throughout the substrate. That is, in some embodiments, no additional substrate or housing element need be positioned between the heat source and the substrate to be heated. Furthermore, because the apparatus of the present disclosure allows the surface of the substrate to directly contact the surface of the heating device (e.g., a thermal cycler), the temperature of the substrate may be more easily controlled by a user and may be heated to a desired temperature in a shorter time than when using an apparatus that does not allow surface contact between the substrate and the heating device (e.g., a thermal cycler). Thus, a sample (e.g., a biological sample) on the substrate can be uniformly and controllably heated.

In one aspect, the present disclosure is directed to a support apparatus comprising: a substrate holder comprising: a cover including opposing first sidewalls, the cover configured to receive a gasket; a base comprising opposing first sidewalls, the base configured to receive a sample substrate; and first and second locking tabs including a movable tab coupled to the first sidewall of one of the cover and the base and a non-movable tab coupled to the first sidewall of the other of the cover and the base, the first locking tab having a length greater than a length of the second locking tab, wherein the first locking tab extends along a length of the liner, and wherein the movable tab is configured to engage the non-movable tab to releasably secure the cover to the base.

In some embodiments, the first locking tab has a height approximately equal to the height of the liner. In some embodiments, the first locking tab has a length approximately equal to the length of the liner. In some embodiments, the device further comprises at least one hinge, and the at least one hinge extends from one of the second sidewalls of the cover to one of the second sidewalls of the base. In some embodiments, at least one hinge is a breakable hinge. In some embodiments, the cover and the base are pivotably connected by at least one hinge.

In some embodiments, the sample substrate rests on top of the base of the substrate holder. In some embodiments, the cover includes a plurality of ribs extending from a surface of the cover, and the liner is positioned in alignment on the surface of the cover by the plurality of ribs. In some embodiments, the gasket is positioned such that when the sample substrate is secured by the substrate holder and the substrate holder is in the closed position, an airtight seal is formed between the gasket and the sample substrate. In some embodiments, the cover comprises at least two apertures.

In some embodiments, the gasket comprises at least two openings, wherein the at least two openings are positioned such that the at least two apertures are aligned with the at least two openings when the sample substrate is secured by the substrate holder and the substrate holder is in the closed position. In some embodiments, the gasket is configured to prevent fluid transfer between the at least two openings when the cover is in the closed position. In some embodiments, the sample substrate is a slide. In some embodiments, the base includes a lip extending around a perimeter of the base, the lip configured to retain the sample substrate within the base. In some embodiments, the cover includes a plurality of ribs extending from a surface of the cover, and the sample substrate engages at least one of the plurality of ribs when the substrate holder is in the closed position. In some embodiments, the base includes an opening that exposes at least a portion of the sample substrate when the sample substrate is placed in the base.

In some embodiments, each of the first sidewalls of the cover is substantially orthogonal to the second sidewall of the cover. In some embodiments, the cover is configured to fold onto the base when the substrate holder is in the closed position. In some embodiments, the length of the first locking tab is at least 90% of the length of the liner, such as at least 75%, 80%, 90%, 100%, 120%, or at least 150% of the length of the liner, about 75% to about 150% of the length of the liner. In some embodiments, the length of the first locking tab relative to the length of the gasket is about 1:0.9 to about 1:1.1, about 1:0.8 to about 1:1.2, about 1:0.5 to about 1: 1.5.

In another aspect, the present disclosure relates to a support apparatus comprising: a substrate holder comprising: a cover comprising opposing first and second sidewalls, the cover configured to receive the gasket; a base comprising opposing first and second sidewalls, the base configured to receive a sample substrate, each of the first sidewalls of the base and cover being longer than each of the second sidewalls of the base and cover; and a locking tab including a movable tab and a non-movable tab, one of the movable tab and the non-movable tab coupled to one of the first sidewalls of the cover and the other of the movable tab and the non-movable tab coupled to one of the first sidewalls of the base, the locking tab having a length approximately equal to a length of the liner, wherein the locking tab extends along the length of the liner.

In some embodiments, the sample substrate rests on top of the base of the substrate holder. In some embodiments, the cover includes a plurality of ribs extending from a surface of the cover, and the liner is positioned in alignment on the surface of the cover by the plurality of ribs. In some embodiments, the gasket is positioned such that when the sample substrate is secured by the substrate holder and the substrate holder is in the closed position, an airtight seal is formed between the gasket and the sample substrate. In some embodiments, the cover comprises at least two apertures. In some embodiments, the gasket comprises at least two openings, wherein the at least two openings are positioned such that the at least two apertures are aligned with the at least two openings when the sample substrate is secured by the substrate holder and the substrate holder is in the closed position. In some embodiments, the gasket is configured to prevent fluid transfer between the at least two openings when the substrate holder is in the closed position.

In some embodiments, the sample substrate comprises a slide. In some embodiments, the base includes a lip extending around a perimeter of the base, the lip configured to retain the sample substrate within the base. In some embodiments, the cover comprises a plurality of ribs extending from a surface of the cover, and the sample substrate engages at least one of the plurality of ribs when the substrate holder is in the closed position. In some embodiments, the base includes an opening that exposes at least a portion of the sample substrate when the sample substrate is placed in the base. In some embodiments, the first sidewall of the cover is substantially orthogonal to the second sidewall of the cover. In some embodiments, the support device further comprises at least one breakable hinge extending from one of the second side walls of the cover to one of the second side walls of the base, such that when the substrate holder is in the closed position, the cover is configured to fold onto the base to secure the sample substrate and the pad between the cover and the base. In some embodiments, the cover and the base are pivotably connected by at least one hinge extending from one of the second sidewalls of the cover to one of the second sidewalls of the base, such that when the substrate holder is in the closed position, the cover is configured to fold over onto the base to secure the sample substrate and the pad between the cover and the base. In some embodiments, the cover and the base are pivotably connected by at least one breakable hinge.

In another aspect, the present disclosure is directed to a method of incubating a sample disposed on a sample area of a substrate, the method comprising: mounting a substrate on a base of a substrate holder of a support apparatus, the substrate comprising a sample; moving a cover of the substrate holder toward a base of the substrate holder such that first and second movable tabs on one of the base and the cover engage first and second non-movable tabs on the other of the base and the cover, the first movable tab having a length greater than a length of the second movable tab and extending along a length of a pad received by the cover; positioning the substrate and the support device on a heating apparatus; and activating the heating device to transfer heat to the sample.

In some embodiments, the substrate comprises a glass slide. In some embodiments, at least 60% of the sample area is covered by the support device when the substrate holder is coupled to the support device. In some embodiments, the first movable tab has a height approximately equal to the height of the cushion. In some embodiments, the first movable tab has a length approximately equal to the length of the cushion.

All publications, patents, patent applications, and information available on the internet and mentioned in this specification are herein incorporated in their entirety by reference to the same extent as if each individual publication, patent application, or information item was specifically and individually indicated to be incorporated by reference. If publications, patents, patent applications, and information items incorporated by reference conflict with the disclosure contained in the specification, the specification is intended to supersede and/or take precedence over any such conflicting material.

Where values are described in ranges, it is understood that the description includes disclosure of all possible sub-ranges within such ranges, as well as specific values falling within such ranges, whether or not the specific value or the specific sub-range is explicitly recited.

The term "each," when used to refer to a set of items, is intended to identify a single item in the set, but does not necessarily refer to each item in the set, unless specifically stated otherwise, or unless the context of use clearly dictates otherwise.

Various embodiments of features of the present disclosure are described herein. It should be understood, however, that these embodiments are provided by way of example only, and that numerous modifications, changes, and substitutions will occur to those skilled in the art without departing from the scope of the present disclosure. It should also be understood that various alternatives to the specific embodiments described herein are also within the scope of the disclosure.

Drawings

The following drawings illustrate certain embodiments of the features and advantages of the present disclosure. These examples are not intended to limit the scope of the appended claims in any way. In the drawings, like numbering represents like elements.

FIG. 1 is a perspective view of an exemplary support device in a closed position.

Fig. 2 is a perspective view of the example support apparatus of fig. 1 in an open position.

Fig. 3 is an exploded view of the exemplary support device of fig. 1.

Fig. 4 is a perspective view of the exemplary support device and base of fig. 1.

FIG. 5 is a perspective view of an exemplary support apparatus in a closed position.

Detailed Description

I. Introduction to

The present disclosure describes devices and methods for spatial analysis of biological samples. This section describes certain general terms, analytes, sample types, and preparation steps referred to in subsequent sections of this disclosure.

1. Spatial analysis

The tissue and cells may be obtained from any source. For example, tissues and cells may be obtained from single or multi-cellular organisms (e.g., mammals). Tissues and cells obtained from mammals, such as humans, often have different analyte levels (e.g., gene and/or protein expression) which can lead to differences in cell morphology and/or function. The location of cells or subsets of cells (e.g., neighboring cells and/or non-neighboring cells) within a tissue can affect, for example, the fate, behavior, morphology, and signaling of the cells as well as cross-talk with other cells in the tissue. Information about differences in the levels (gene and/or protein expression) of different intracellular analytes in mammalian tissues may also help physicians select or implement effective therapies, and may allow researchers to identify and elucidate cellular morphology and/or cellular function in single-or multi-cellular organisms (e.g., mammals) based on the detected differences in the levels of different intracellular analytes in tissues. Differences in the levels of analytes within different cells in mammalian tissue can also provide information on how tissues (e.g., healthy and diseased tissues) function and/or develop. Differences in the levels of the analyte in different cells of mammalian tissue may also provide information on the different mechanisms of disease pathogenesis in the tissue and the mechanism of action of therapeutic treatment in the tissue. Differences in the levels of analytes within different cells in mammalian tissue can also provide information about the mechanism of drug resistance and its development in mammalian tissue. The difference in the presence or absence of an analyte within different cells in a multicellular organism (e.g., mammalian) tissue can provide information about the mechanism of resistance and its development in the multicellular organism tissue.

The support devices provided herein can be used with spatial analysis methods that provide high spatial resolution of mass analyte levels and/or expression data for multiple analytes within a sample, e.g., while maintaining a natural spatial background. Spatial analysis methods include, for example, the use of capture probes that include a spatial barcode (e.g., a nucleic acid sequence that provides information about the location of the capture probe on a substrate, which location is associated with a location within a cell or tissue sample (e.g., a mammalian cell or mammalian tissue sample)) and a capture domain that is capable of binding to an analyte (e.g., a protein and/or a nucleic acid) produced by and/or present in the cell or tissue.

Binding of the analyte to the capture probe can be detected using a variety of different methods, such as nucleic acid sequencing, fluorophore detection, nucleic acid amplification, nucleic acid ligation detection, and/or nucleic acid cleavage product detection. In some examples, detection is used to correlate and correlate a particular spatial barcode with a particular analyte produced and/or present at a particular location in a cell or tissue (e.g., a mammalian cell or tissue).

The capture probes may, for example, be attached to a surface, such as a solid array, a bead (bead), or a cover slip. In some examples, the capture probes are not attached to the surface. In some examples, the capture probes can be encapsulated, embedded within, or layered on a surface of a permeable composition (e.g., any of the substrates described herein). For example, the capture probes may be encapsulated or disposed within permeable beads (e.g., gel beads). In some examples, the capture probes may be encapsulated, embedded within, or layered on a surface of a substrate (e.g., any of the exemplary substrates described herein, such as a hydrogel or porous membrane). When the capture probes are attached to a substrate, such as the surface of a slide, the attachment can be direct or indirect (e.g., via a link).

In some examples, cells or tissue samples comprising cells are contacted with capture probes attached to a substrate (e.g., a surface of a substrate), and the cells or tissue samples are permeabilized to allow analytes to be released from the cells and intermixed with the capture probes attached to the substrate. In some examples, analytes released from the cells passively migrate to the substrate to intermix with the capture probes (e.g., via gravity). In some examples, analytes released from the cells may be actively directed to capture probes attached to a substrate using a variety of methods, such as electrophoresis, chemical gradients, pressure gradients, fluid flow, or magnetic fields.

Non-limiting aspects of the support device are described in PCT/US2019/065100, which is incorporated herein by reference in its entirety and may be used herein in any combination. Other non-limiting aspects of the support apparatus are described herein.

(a)General terms

(i) Biological sample

The devices and methods described in the present disclosure may be used to support a substrate configured to receive one or more biological samples. As used herein, a "biological sample" is a sample that may be obtained from a subject for analysis using any of a variety of techniques, including but not limited to biopsy, surgery, and Laser Capture Microscopy (LCM), and typically includes cells and/or other biological material from the subject, such as suspended biological material or as a tissue section from a tissue sample. In addition to the test described below, the biological sample can be obtained from a non-mammalian organism (e.g., a plant, an insect, an arachnid, a nematode (e.g., caenorhabditis elegans), a fungus, an amphibian, or a fish (e.g., zebrafish)). Biological samples can be obtained from: prokaryotes, such as bacteria, e.g., Escherichia coli, Staphylococcus, or Mycoplasma pneumoniae; archaea; viruses such as hepatitis c virus or human immunodeficiency virus; or a viroid. The biological sample may be obtained from a eukaryote, such as a Patient Derived Organoid (PDO) or a Patient Derived Xenograft (PDX). Biological samples may include organoids, which are miniaturized and simplified versions of organs produced in vitro that can show realistic microdissection in three dimensions. Organoids can be generated from one or more cells of a tissue, embryonic stem cells, and/or induced pluripotent stem cells, which can self-organize in three-dimensional culture due to their ability to self-renew and differentiate. In some embodiments, the organoid is a brain organoid, an intestinal organoid, a stomach organoid, a tongue organoid, a thyroid organoid, a thymus organoid, a testis organoid, a liver organoid, a pancreas organoid, an epithelial organoid, a lung organoid, a kidney organoid, a gastral embryo, a heart organoid, or a retinal organoid. The subject from which the biological sample can be obtained can be a healthy or asymptomatic individual, an individual having or suspected of having a disease (e.g., cancer) or of being predisposed to a disease, and/or an individual in need of treatment or suspected of being in need of treatment.

The biological sample may be from a homogeneous culture or population of the subject or organism referred to herein, or from a collection of several different organisms, for example in a community or ecosystem.

The biological sample may include one or more diseased cells. The diseased cells may have altered metabolic characteristics, gene expression, protein expression, and/or morphological features. Examples of diseases include inflammatory diseases, metabolic diseases, neurological diseases, and cancer. Cancer cells can be obtained from solid tumors, hematologic malignancies, cell lines, or as circulating tumor cells.

The biological sample may also include fetal cells. For example, a procedure such as amniocentesis may be performed to obtain a fetal cell sample from the maternal circulation. The sequence of the fetal cell can be used to identify any of a variety of genetic diseases, including, for example, aneuploidy, such as down's syndrome, edwards syndrome, and patau syndrome. Further, the cell surface characteristics of the fetal cells can be used to identify any of a variety of conditions or diseases.

The biological sample may also include immune cells. Sequence analysis of the immune repertoire of these cells, including genomic, proteomic, and cell surface features, can provide rich information to aid in understanding the state and function of the immune system. For example, determination of Minimal Residual Disease (MRD) status (e.g., negative or positive) in Multiple Myeloma (MM) patients following autologous stem cell transplantation is considered a predictor of MRD in MM patients (see, e.g., U.S. patent application publication No. 2018/0156784, which is incorporated herein by reference in its entirety).

Examples of immune cells in a biological sample include, but are not limited to, B cells, T cells (e.g., cytotoxic T cells, natural killer T cells, regulatory T cells, and T helper cells), natural killer cells, cytokine-induced killer (CIK) cells, myeloid cells such as granulocytes (basophils, eosinophils, neutrophils/super-segmental neutrophils), monocytes/macrophages, mast cells, platelets/megakaryocytes, and dendritic cells.

Biological samples may include any number of macromolecules, such as cellular macromolecules and organelles (e.g., mitochondria and nuclei). The biological sample may be a nucleic acid sample and/or a protein sample. The biological sample may be a carbohydrate sample or a lipid sample. The biological sample may be obtained as a tissue sample, such as a tissue section, biopsy, core biopsy, needle aspirate or fine needle aspirate. The sample may be a fluid sample, such as a blood sample, a urine sample or a saliva sample. The sample may be a skin sample, a colon sample, a cheek swab, a histological sample, a histopathological sample, a plasma or serum sample, a tumor sample, a living cell, a cultured cell, a clinical sample such as a whole blood product or blood derived product, a blood cell or a cultured tissue or cell, including a cell suspension.

The cell-free biological sample may comprise extracellular polynucleotides. Extracellular polynucleotides can be isolated from body samples such as blood, plasma, serum, urine, saliva, mucosal excretions, sputum, feces, and tears. The biological sample may include an analyte of interest, or more than one analyte of interest.

(ii) Is tested

As used herein, the term "subject" is an animal such as a mammal (e.g., a human or non-human ape) or an avian (e.g., an avian), or other organism such as a plant. Examples of subjects include, but are not limited to, mammals such as rodents, mice, rats, rabbits, guinea pigs, ungulates, horses, sheep, pigs, goats, cows, cats, dogs, primates (i.e., human or non-human primates); plants, such as arabidopsis, maize, sorghum, oats, wheat, rice, canola or soybean; algae such as chlamydomonas reinhardtii; nematodes, such as caenorhabditis elegans; insects such as drosophila melanogaster, mosquitoes, fruit flies, or bees; arachnids, such as spiders; fish, such as zebrafish; a reptile; amphibians, such as frogs or xenopus; dictyostelium discodermatum; fungi, such as pneumocystis carinii, fugu rubripes, yeast, saccharomyces cerevisiae or schizosaccharomyces pombe; or plasmodium falciparum.

(iii) Substrate attachment

In some embodiments, the biological sample may be attached to a substrate. Examples of substrates suitable for this purpose are described in detail below. Attachment of the biological sample may be irreversible or reversible, depending on the nature of the sample and subsequent steps in the assay method.

In certain embodiments, the sample may be reversibly attached to the substrate by applying a suitable polymeric coating to the substrate and contacting the sample with the polymeric coating. The sample can then be separated from the substrate using an organic solvent that at least partially dissolves the polymer coating. Hydrogels are examples of polymers suitable for this purpose.

More generally, in some embodiments, the substrate may be coated or functionalized with one or more substances to facilitate attachment of the sample to the substrate. Suitable substances that may be used to coat or functionalize the substrate include, but are not limited to, lectins, polylysines, antibodies, and polysaccharides.

(iv) Substrate

For analytical methods that use a substrate (e.g., spatial array-based analytical methods), the substrate functions as a support to attach the capture probes directly or indirectly to the array features. Further, in some embodiments, substrates (e.g., the same substrate or different substrates) can be used to provide support to a biological sample, particularly, for example, a thin tissue section. Thus, as used herein, a "substrate" is a support that is insoluble in aqueous liquids and allows positioning of biological samples, analytes, features and/or capture probes on the substrate.

Further, as used herein, "substrate," when the modifier "chemical" is not used above, refers to a member having at least one surface that generally functions to provide physical support for a biological sample, an analyte, and/or any other chemical and/or physical moiety, reagent, and structure that may be used in various analytical methods. The substrate can be made from a variety of solid materials, gel-based primers, colloidal materials, semi-solid materials (e.g., at least partially cross-linked materials), fully or partially cured materials, and materials that undergo a phase change or transformation to provide physical support. Examples of substrates that may be used in the methods and devices described herein include, but are not limited to, slides (e.g., slides formed from various glasses, slides formed from various polymers), hydrogels, layers and/or films, membranes (e.g., porous membranes), wafers, plates, or combinations thereof. In some embodiments, as will be discussed in further detail below, the substrate can optionally include functional elements, such as grooves, protruding structures, microfluidic elements (e.g., channels, reservoirs, electrodes, valves, seals), and various indicia.

1) Substrate Properties

The substrate may generally have any suitable form or format that can be accommodated by the devices disclosed herein. For example, the substrate may be flat, curved, e.g., convexly or concavely curved towards a section where interaction between the biological sample (e.g., tissue sample) and the substrate occurs. In some embodiments, the substrate is flat, such as a flat surface, a chip, or a slide. The substrate may include one or more patterned surfaces (e.g., channels, wells, protrusions, ridges, divots, etc.) within the substrate.

The substrate may be of any desired shape. For example, the substrate may typically be a thin, flat shape (e.g., square or rectangular). In some embodiments, the base structure has rounded corners (e.g., for added safety or robustness). In some embodiments, the base structure has one or more truncated corners (e.g., for use with a slide holder or cross table). In some embodiments, when the base structure is planar, the base structure may be any suitable type of support (e.g., a chip or slide, such as a microscope slide) having a planar surface.

The substrate can optionally include various structures such as, but not limited to, protrusions, ridges, and channels. The substrate may be micropatterned to limit lateral diffusion (e.g., to prevent spatial bar code overlap). Substrates modified with such structures may be modified to allow association of analytes, features (e.g., specific locations where barcodes are deposited, beads), or probes at individual sites. For example, the locations where the substrate is modified with various structures may be continuous with other locations (e.g., when the device is in the closed position, the locations may be located within a section of the substrate enclosed by one of the gasket openings) or discontinuous (e.g., when the support device is in the closed position, a first location may be located within a first section of the substrate enclosed by a first gasket opening and a second location may be located within a second section of the substrate enclosed by a second gasket).

In some embodiments, the surface of the substrate may be modified such that discrete sites are formed that may only have or accommodate a single feature. In some embodiments, the surface of the substrate may be modified such that the features are located at random locations (e.g., random locations within the section of the substrate enclosed by the gasket opening when the support device is in the closed position).

In some embodiments, the surface of the substrate is modified to contain one or more wells using techniques such as (but not limited to) stamping, microetching, or molding techniques. In some embodiments where the substrate comprises one or more wells, the substrate may be a concave slide or a cavity slide. For example, the wells may be formed by one or more shallow depressions on the surface of the substrate. In some embodiments, when the substrate comprises one or more wells, the wells can be formed by attaching a cartridge (e.g., a cartridge containing one or more chambers) to a surface of the substrate structure.

In some embodiments, the structures (e.g., wells or features) of the substrate may each carry one or more different capture probes. The different capture probes attached to each structure may be identified according to the position of the structure in or on the surface of the substrate. Exemplary substrates include arrays in which individual structures are located on a substrate, including, for example, those having wells or locations on the substrate that receive features.

In some embodiments where the substrate is modified to contain one or more structures, including but not limited to wells, protrusions, ridges, features, or markings, the structures may include physically altered sites. For example, substrates modified with various structures may include physical properties including, but not limited to, physical configurations, magnetic or compressive forces, chemical functionalization sites, chemical alteration sites, and/or electrostatic alteration sites. In some embodiments where the substrate is modified to contain various structures, including but not limited to wells, protrusions, ridges, features, or markings, the structures are applied in a pattern. Alternatively, the structures may be randomly distributed.

The substrate (e.g., or beads or features on an array) can include tens to hundreds of thousands or millions of individual oligonucleotide molecules (e.g., at least about 10,000, 50,000, 100,000, 500,000, 1,000,000, 10,000,000, 100,000,000, 1,000,000, or 10,000,000,000 oligonucleotide molecules).

In some embodiments, the substrate comprises one or more markings on the substrate surface, e.g., to provide guidance for correlating spatial information with a characterization of an analyte of interest. For example, the substrate may be marked with a wire grid (e.g., to allow easy estimation of the size of the object seen under magnification and/or to provide reference sections for counting objects). In some embodiments, fiducial marks may be included on the substrate. Such markings may be made using techniques including, but not limited to, printing, sandblasting, and deposition on the surface.

A variety of different substrates may be used for this purpose. In general, the substrate can be any suitable support material that can be accommodated by the disclosed apparatus. Exemplary substrates include, but are not limited to, glass, modified and/or functionalized glass, hydrogels, films, membranes, plastics (including, for example, acrylics, polystyrene, copolymers of styrene and other materials, polypropylene, polyethylene, polybutylene, polyurethane, teflon^TMCyclic olefin, polyimide, etc.), nylon, ceramic, resin, japanese pulsatilla (Zeonor), silica or silica substrate including silicon and modified silicon, carbon, metal, inorganic glass, optical fiber bundle, and polymers such as polystyrene, Cyclic Olefin Copolymer (COC), Cyclic Olefin Polymer (COP), polypropylene, polyethylene carbonate, or combinations thereof.

In the example of the substrate material discussed above, polystyrene is a hydrophobic material suitable for binding negatively charged macromolecules because it typically contains few hydrophilic groups. For nucleic acids immobilized on a slide glass, the immobilization of nucleic acids can be increased by increasing the hydrophobicity of the glass surface. Such enhancements may allow for relatively more tightly packed formations (e.g., providing improved specificity and resolution).

2) Conductive substrate

In some embodiments, the substrate may be a conductive substrate. Conductive substrates (e.g., electrophoretically compatible arrays) produced as described herein can be used for spatial detection of analytes.

In some embodiments, the conductive substrate may comprise glass (e.g., a glass slide) that has been coated with a substance or otherwise modified to impart conductive properties to the glass. In some embodiments, the slide can be coated with a conductive coating. In some embodiments, the conductive coating comprises Tin Oxide (TO) or Indium Tin Oxide (ITO). In some embodiments, the conductive coating comprises a Transparent Conductive Oxide (TCO). In some embodiments, the conductive coating comprises aluminum-doped zinc oxide (AZO). In some embodiments, the conductive coating comprises fluorine-doped tin oxide (FTO).

In some embodiments, arrays spotted or printed with oligonucleotides (e.g., capture probes) can be produced on a conductive substrate (e.g., any of the conductive substrates described herein). For example, the arrays described herein may be compatible with active analyte capture methods (e.g., including, but not limited to, electrophoretic capture methods). In some embodiments, the conductive substrate is a porous medium. Non-limiting examples of porous media that can be used in methods employing active analyte capture include nitrocellulose or nylon membranes. In some embodiments, the porous medium that may be used in the methods employing active analyte capture described herein comprises paper. In some embodiments, the oligonucleotides may be printed on a paper substrate. In some embodiments, the printed oligonucleotides can interact with the substrate (e.g., interact with the fibers of the paper). In some embodiments, the printed oligonucleotides may be covalently bound to a substrate (e.g., to the fibers of paper). In some embodiments, the oligonucleotides in the molecular precursor solution can be printed on a conductive substrate (e.g., paper). In some embodiments, the molecular precursor solution may be polymerized, thereby creating a gel pad on a conductive substrate (e.g., paper). In some embodiments, the molecular precursor solution may be cured by photopolymerization (e.g., photocuring). In some embodiments, gel beads containing oligonucleotides (e.g., barcoded oligonucleotides, such as capture probes) can be printed on a conductive substrate (e.g., paper). In some embodiments, the printed oligonucleotides may be covalently attached into the gel matrix.

3) Coating layer

In some embodiments, the surface of the substrate may be coated with a cell permissive coating to allow for the adhesion of living cells. A "cell permissive coating" is a coating that allows or aids cells to maintain cell viability (e.g., remain viable) on a substrate. For example, the cell permissive coating may enhance cell attachment, cell growth, and/or cell differentiation, e.g., the cell permissive coating may provide nutrients to living cells. The cell permissive coating may include a biological material and/or a synthetic material. Non-limiting examples of cell permissive coatings include coatings formed from one or more extracellular matrix (ECM) components (e.g., proteoglycans and fibrous proteins such as collagen, elastin, fibronectin, and laminin), polylysine, poly (L) -ornithine, and/or biocompatible silicones (e.g.,

) Is a characterized coating. For example, a cell permissive coating comprising one or more extracellular matrix components may comprise type I collagen, type II collagen, type IV collagen, elastin, fibrin, laminin, and/or fiberglass protein. In some embodiments, the cell permissive coating comprises a solubilized basement membrane formulation (e.g., extracted from Engelbreth-Holm-swarm (EHS) mouse sarcoma

). In some embodiments, the cell permissive coating comprises collagen. Thin and thinThe cell permissive coating can be used to culture adherent cells on the spatial barcode array, or to maintain cell viability of a tissue sample or section when contacted with the spatial barcode array.

In some embodiments, the substrate is coated with a surface treatment agent, such as poly (L) -lysine. Additionally or alternatively, the substrate may be treated by silanization, for example, with epoxy silanes, amino silanes, and/or with polyacrylamides.

In some embodiments, the substrate is treated to minimize or reduce non-specific analyte contamination within or between features. For example, the treatment may include coating the substrate with a hydrogel, film, and/or membrane that forms a physical barrier to non-specific hybridization. Any suitable hydrogel may be used. For example, hydrogel matrices prepared according to the methods set forth in U.S. patent nos. 6,391,937, 9,512,422, and 9,889,422 and U.S. patent application publications nos. u.s.2017/0253918 and u.s.2018/0052081 may be used. Each of the foregoing is incorporated by reference herein in its entirety.

The treatment can include adding a reactive or activatable functional group that becomes reactive upon application of a stimulus (e.g., a photoreactive functional group). The treatment may include treatment with a polymer having one or more physical properties (e.g., mechanical, electrical, magnetic, and/or thermal) to minimize non-specific binding (e.g., activating the substrate at certain locations to allow analyte hybridization at those locations).

By "removable coating" is meant a coating that is removable from the surface of the substrate after application of the release agent. In some embodiments, the removable coating comprises a hydrogel as described herein, e.g., a hydrogel comprising a polypeptide-based primer. Non-limiting examples of hydrogels with polypeptide-based primers include synthetic peptide-based primers characterized by a combination of spider silk and transmembrane segments of human muscle L-type calcium channels (e.g.,

) A pharmaceutical composition comprises a repetitive arginine-alanine-aspartate-alanine sequence (RADARA)DARADA) amphiphilic 16 residue peptides (e.g.,

) EAK16(AEAEAKAKAEAEAKAK), KLD12(KLDLKLDLKLDL) and PGMATRIX^TM。

In some embodiments, the hydrogel in the removable coating is a stimuli-reactive hydrogel. The stimulus-responsive hydrogel can undergo a gel-to-solution and/or gel-to-solid transition upon application of one or more external triggers (e.g., release agents). See Willner Acc. chem. Res. (chemical research statement) 50: 657-. Non-limiting examples of stimulus-reactive hydrogels include thermally reactive hydrogels, pH reactive hydrogels, photoreactive hydrogels, redox reactive hydrogels, analyte reactive hydrogels, or combinations thereof. In some embodiments, the stimuli-reactive hydrogel can be a multi-stimulus-reactive hydrogel.

By "release agent" or "external trigger" is meant a formulation that allows the removable coating to be removed from the substrate when the release agent is applied to the removable coating. External triggers or release agents may include physical triggers such as thermal, magnetic, ultrasonic, electrochemical and/or light stimuli, and chemical triggers such as pH, redox reactions, supramolecular complexes and/or biocatalytically driven reactions. See, for example, Gels (2018),4454: doi:10.3390/Gels4020054 to Echeverria et al, which is incorporated herein by reference in its entirety. The type of "release agent" or "external trigger" may depend on the type of removable coating. For example, the removable coating with the redox-reactive hydrogel can be removed upon application of a release agent including a reducing agent such as Dithiothreitol (DTT). As another example, the pH-reactive hydrogel may be removed upon application of a release agent that changes the pH. In some embodiments, the biological sample may be confined to a particular region or segment. For example, the biological sample may be secured to a slide and a chamber, pad, or cage positioned over the biological sample to serve as a containment area or frame in which the biological sample is deposited.

Sequence ID number: 7GGTGACTCTAGATAACCT

2.Additional support device embodiments

In some embodiments, the support device can be part of a system for heating a substrate (e.g., system 3102 as described in PCT/US2019/065100, which is incorporated herein by reference in its entirety), which can also include a plate. The plate may be configured to be received by a heating device (e.g., a thermal cycler) and provide heat transfer between the heating device and the support device. A support device (e.g., substrate holder 3150 as described in PCT/US 2019/065100) can hold one or more substrates (e.g., one or more slides) and can be removably coupled to the plate to facilitate heat transfer from the plate to the one or more substrates. The support device may include a bottom member and a top member. In some embodiments, the base device may further comprise a slide. In some embodiments, the support device may include a liner positioned inside the support device. In some embodiments, the support device may include an engagement mechanism (e.g., a screw) for coupling the bottom member and the top member.

In some embodiments, the support means may be a one-piece component (e.g., substrate holder 4400 as described in PCT/US 2019/065100) that receives the gasket and the substrate. In some embodiments, the support means may include one or more fasteners (such as side-mounted pressure latches 4410 as described in PCT/US 2019/065100) for snap engagement of the base. In some embodiments, the support device may further comprise one or more tabs (e.g., a first tab 4412a and a second tab 4412b as described in PCT/US 2019/065100) configured to engage the substrate. The support device may include a bottom surface defining a plurality of apertures configured to align with the plurality of apertures of the liner.

In some embodiments, the support means may comprise a top part and a bottom part connected via one or more hinges (e.g. hinge 7360 as described in PCT/US 2019/065100) extending from the side walls of the bottom part. The support means may further comprise one or more engagement features (e.g. first and second notches 7358a, 7358b as described in PCT/US 2019/06510) projecting from the side wall of the top part, which features are configured to engage with one or more tabs projecting from the side wall of the bottom part, thereby effecting closure of the support means.

II. supporting device

Embodiments may provide one or more of the following advantages.

The devices provided herein can be used to ensure that the temperature of the substrate and any sample and/or reagents supported on the surface of the substrate is uniformly and consistently controlled during a sample preparation and/or analysis protocol. During such protocols, uneven heating can lead to sample preparation failures. Further, even where the sample is heated relatively uniformly, condensate contacting the sample may damage certain reactions that are part of the protocol or otherwise affect the chemical reactions that occur. In various embodiments, the apparatus provides heating of multiple surfaces of a substrate (e.g., in a slide cassette or substrate holder), and may include features that facilitate heat transfer from the heating element to the substrate and that may reduce or prevent condensate formation in certain regions of the substrate (e.g., in sample wells or on regions of the substrate surface). In addition, the devices provided herein can mitigate cross-contamination of samples and/or reagents from different locations on the substrate. For example, the liner of the device can provide discrete biological sample sections and a vapor barrier from one well to the next. Further, the liner may impede the flow of reagents from one well to another. In some embodiments, the devices provided herein include a lock that helps secure the gasket in place when the support device is in the closed position.

1-4, an embodiment of an exemplary support apparatus can include a substrate and a substrate holder. The substrate holder may include a pad, a cover, and a base. In some embodiments, the cover and base are integrally connected (e.g., the substrate holder is a one-piece design). In some embodiments, the substrate holder is manufactured using injection molding techniques. Non-limiting materials for making the support devices of the present disclosure include polypropylene homopolymers. In some embodiments, the substrate holder is disposable. In some embodiments, the substrate holder is reusable.

In some embodiments, the substrate holder receives a substrate, such as a slide, for example a glass slide. In some embodiments, the substrate holder includes an attachment mechanism for coupling and/or securing the substrate to the substrate holder. The cover may be configured to receive the gasket, or may be co-molded with the gasket. The cover may include a plurality of ribs extending from a surface of the cover. Further, the base may be configured to receive a substrate. In some examples, the support device can further include at least one pair of locking tabs, wherein each locking tab includes a movable tab coupled to the first sidewall of the cover and a non-movable tab coupled to the first sidewall of the base. In other examples, each locking tab includes a movable tab coupled to the first sidewall of the base and a non-movable tab coupled to the first sidewall of the cover. The movable tab may be configured to engage with the non-movable tab to releasably secure the cover to the base. In some embodiments, a length of a first locking tab of the pair of locking tabs is greater than a length of a second locking tab of the pair of locking tabs.

In some examples, the cover and the base are pivotably connected together by at least one hinge. In some embodiments, the hinge may be a living hinge. In some embodiments, the cover and the base are pivotally connected together by two or more hinges. In some embodiments, the at least one hinge extends from the second sidewall of the cover to the second sidewall of the base. For example, the cover may be configured to fold onto the base to secure the gasket and the substrate between the cover and the base when the substrate holder is in the closed position. In some embodiments, the pivotable connection may be broken at any stage, such that there is no longer a pivotable connection between the cover and the base.

Further, the support device can include a substrate (e.g., a slide) configured to receive the sample. In some embodiments, the sample may be a biological sample. In some embodiments, the sample may be any biological sample defined elsewhere in the present disclosure. The substrate includes a first surface and a second surface. In some examples, the first surface of the substrate is configured to receive a sample. In some embodiments, the substrate comprises a sample region. In some examples, the sample region receives one or more samples.

Fig. 1 shows the support device 100 in a closed position. The support device 100 includes a substrate holder 101 and a substrate 110. Specifically, fig. 1 shows a substrate holder 101 having a base 134 and a cover 136. The base 134 and cover 136 have a first pair of sidewalls 103 and a second pair of sidewalls 105. Each of the first sidewalls 103 may be longer than each of the second sidewalls 105. Thus, the base 134 and the cover 136 have a substantially rectangular shape. In some embodiments, the base 134 and the cover 136 have a circular, square, triangular, or any other suitable shape. In some embodiments, the first sidewall 103 of the cover 136 is substantially orthogonal to the second sidewall 105 of the cover 136. For example, the first sidewall 103 and the second sidewall 105 of the cover 136 may form an angle between 85 and 95 degrees. In some embodiments, the first sidewall 103 of the base 134 is substantially orthogonal to the second sidewall 105 of the base 134. For example, the first sidewall 103 and the second sidewall 105 of the base 134 may form an angle between 85 and 95 degrees. The cover 136 includes a top surface 102 through which a plurality of apertures 104 are defined. In some embodiments, the plurality of apertures 104 may be defined such that when the gasket in the closed position is positioned between the cover 136 and the base 134, the plurality of apertures are aligned with the plurality of openings of the gasket. In some embodiments, the plurality of apertures 104 can be defined such that the plurality of apertures are aligned with a sample area (e.g., slide) of the substrate when the support device is in the closed position. In some embodiments, the plurality of wells 104 can provide a user with access to a substrate (e.g., a sample area on a substrate) to view a sample on the sample area and/or deliver a solution directly onto the surface of the sample or substrate on the sample area, for example, when the support device is in a closed position.

The substrate holder 101 may include at least a pair of locking tabs 107 configured to releasably secure, close, lock, secure, and/or engage the base 134 with the cover 136. Each locking tab 107 may include a movable tab 106 coupled to the first sidewall 103 of the cover 136 and a non-movable tab 108 coupled to the first sidewall 103 of the base 134. In some embodiments, the movable tab 106 is configured to engage with the non-movable tab 108 to releasably secure the cover 136 to the base 134. In alternative examples, the movable tab 106 may be coupled to the second sidewall 105 of the cover 136 and the non-movable tab 108 may be coupled to the second sidewall 105 of the base 134. In some embodiments, one or more locking tabs 107 may be coupled to one or both of first side walls 103, and/or one or more locking tabs 107 may be coupled to one or both of second side walls 105. In some embodiments, the movable tab 106 and the cover 136 are integrally joined. For example, the movable tab 106 and the cover 136 may be formed as a single component during a molding, e.g., injection molding, process. In some embodiments, non-movable tab 108 and base 134 are integrally joined. For example, the non-movable tab 108 and the base 134 may be formed as a single component during molding, such as in an injection molding process. In some examples, the substrate holder 101 may include a plurality of locking tabs 107 (e.g., at least 2, 3, 4, or 5 pairs of locking tabs 107). In some embodiments, the substrate holder 101 includes one locking tab. In some embodiments, any type of fastener that allows the base 134 to releasably engage with the cover 136 may be used, such as, for example, a magnetic fastener, a snap, a hook and loop fastener, a push latch, a screw, a push connector (e.g., a lever, a clip, or a clamp), or any combination thereof. In some embodiments, the substrate holder 101 may also include one or more spring-loaded fasteners.

The substrate holder 101 may include a locking tab 158 configured to releasably secure, close, lock, secure, and/or engage the base 134 with the cover 136. The locking tabs 158 may include a movable tab 162 coupled to the first sidewall 103 of the cover 136 and a non-movable tab 160 coupled to the first sidewall 103 of the base 134. In some embodiments, the movable tab 162 is configured to engage with the non-movable tab 160 to releasably secure the cover 136 to the base 134. In alternative examples, the movable tab 162 may be coupled to the second sidewall 105 of the cover 136 and the non-movable tab 160 may be coupled to the second sidewall 105 of the base 134. In some embodiments, one or more locking tabs 158 may be coupled to one or both of the first side walls 103, and/or one or more locking tabs 158 may be coupled to one or both of the second side walls 105. In some embodiments, the movable tab 162 and the cover 136 are integrally joined. In some embodiments, the non-movable tab 160 and the base 134 are integrally joined. In some examples, the substrate holder 101 may include a plurality of locking tabs 158 (e.g., at least 2, 3, 4, or 5 locking tabs 158). In some embodiments, the substrate holder 101 includes one locking tab 158 and one locking tab 107. In some embodiments, the substrate holder 101 includes one locking tab 158 and a pair of locking tabs 107. In some embodiments, the substrate holder 101 includes one locking tab 158 and three locking tabs 107. In some embodiments, the substrate holder 101 includes one locking tab 158 and four locking tabs 107.

As shown in fig. 1, the surface of the substrate 110 is exposed through the plurality of holes 104 of the substrate holder 101. A plurality of apertures 104 are defined by the top surface 102 of the cover 136. The plurality of apertures 104 are aligned with the plurality of openings of the liner when the support 100 is in the closed position.

Fig. 2 is a perspective view of the substrate holder 101 of fig. 1 in an open position. Substrate holder 201 includes a locking tab 258 comprising a locking tab having a length of l_mAnd a movable tab 262 and having a length l_nNon-movable tab 260. The base retainer 201 includes a pair of locking tabs 207, each locking tab including a length l'_mAnd a movable tab 206 of length l'_nThe non-movable tab 208. The length of locking tab 258 may be greater than the length of the pair of locking tabs 207. For example, the length l of the movable tab 262_mMay be greater than the length l 'of the movable tab 206'_m. In some embodimentsLength l of the movable tab 262_mMay be greater than the length l 'of the movable tab 206'_mThe length is about 40%. In some embodiments, the length lm of the movable tab 262 may be greater than the length l 'of the movable tab 206'_mAt least about 10% to about 80% (e.g., between about 10% to 20%, 20% to 30%, 30% to 40%, 40% to 50%, 50% to 60%, 60% to 70%, or 70% to 80%) long. Length l of non-movable tab 260_nMay be greater than the length l 'of non-movable tab 208'_n. In some embodiments, the length l of the non-movable tab 260_nMay be greater than length l 'of non-movable tab 208'_nThe length is about 40%. In some embodiments, the length l of the non-movable tab 260_nMay be greater than length l 'of non-movable tab 208'_nAt least about 10% to about 80% (e.g., between about 10% to 20%, 20% to 30%, 30% to 40%, 40% to 50%, 50% to 60%, 60% to 70%, or 70% to 80%) long.

In some embodiments, the length l of the movable tab 262_mApproximately equal to the length l of the pad 224_gOr length l of pad 224_g. In some embodiments, the length l of the movable tab 262_mMay be longer than the length l of the pad 224_gThe length is about 5%. In some embodiments, the length l of the movable tab 262_mMay be longer than the length l of the pad 224_gAt least about 5% to about 20% (e.g., between about 5% to 10%, 10% to 15%, 15% to 20%) long. In some embodiments, the length l of the movable tab 262_mAnd length l of pad 224_gAnd (6) aligning. In some embodiments, the length l of the movable tab 262_mIs the length l of the pad 224_gAt least 90%, e.g., length l of pad 224_gAt least 75%, 80%, 90%, 100%, 120%, or at least 150% of pad 224, length l of pad 224_gFrom about 75% to about 150%. In some embodiments, the length l of the movable tab 262_mRelative to the length l of the pad 224_gFrom about 1:0.9 to about 1:1.1, from about 1:0.8 to about 1:1.2, from about 1:0.5 to about 1: 1.5. The length l of the movable tab 262 when the substrate holder 201 is in the closed position_mMay help secure the pad 224 so that it is along the length/of the pad 224_gA force is applied to the gasket wall 209. In some embodiments, the movable tab 262 is centered around the pad 224 (e.g., along the length/of the pad 224)_mCentered). The relatively greater length of locking tab 258 compared to the length of locking tab 207 may facilitate distribution of the force applied to cushion wall 209 over the entire length of cushion 224. In addition, the relatively greater length of the locking tab 258 may make it easier for a user to manipulate the locking tab 258 (e.g., for locking or releasing the locking tab 258).

In fig. 2, the cover 236 and the base 234 are pivotally connected by a pair of hinges 216 that extend from the second sidewall 205 of the cover 236 to the second sidewall 205 of the base 234. In some embodiments, no hinge is present. In some embodiments, the pair of hinges may be a pair of living hinges. In some embodiments, the cover and the base are pivotably connected by at least one hinge. In some embodiments, the one or more hinges may extend from the second sidewall of the cover to the second sidewall of the base. In some embodiments, the cover and base may be pivotally connected by a hinge having a length that spans about 50% or more of the length of the side from which it extends. In some embodiments, the cover and base are pivotally connected together by several hinges. In some embodiments, if a hinge is present, the hinge is breakable such that the cover and the base are separable upon breaking of one or more hinges.

Fig. 2 shows the substrate holder 201 in an open position with the cover 236 in an extended, deployed position away from the base 234. In some embodiments, as shown in fig. 2, the substrate holder 201 is in the open position when the cover 236 is at about 180 ° relative to the base 234. In some embodiments, the substrate holder 201 is in the open position when the cover 236 is at an angle of about 10 ° to about 170 ° relative to the base 234.

The cover 236 has a bottom surface 232 from which a plurality of ribs 218 may extend perpendicularly. In some embodiments, cover 236 is configured to receive pad 224.In some embodiments, the gasket 224 may undergo a compression set of at least about 10%. In some embodiments, the pad 224 may undergo a compression deformation of about 15% or more. In some embodiments, the pad 224 may undergo a compressive deformation of about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 40%, 60%, 70% or more. As used herein, compressive deformation is expressed as a percentage of the original specimen thickness after exposure to a constant compressive force. In some embodiments, the gasket 224 is co-molded with a portion of a substrate holder, such as, for example, the cover 236. In some embodiments, the pad 224 may be subject to a resulting pad height h_gA compressive force varying by about 0.5 millimeters (mm). In some embodiments, the pad 224 may be subject to a resulting pad height h_gA compressive force varying by about 0.1mm, 0.2mm, 0.3mm, 0.4mm, 0.5mm, 0.6mm, 0.7mm, 0.8mm, 0.9mm, 1mm or more. In some embodiments, the gasket 224 is composed of a thermoplastic polymer, such as a thermoplastic elastomer in some embodiments, the gasket 224 is removably adhered to the bottom surface 232 of the cover 236.

The gasket 224 may include a plurality of openings 238. In some embodiments, the plurality of openings 238 may align with the plurality of apertures 104 defined by the surface of the cover 236. For example, in some embodiments, the plurality of openings 238 are positioned such that when the substrate 110 (shown in fig. 1) rests on top of the substrate holder 201 and the substrate holder 201 is in the closed position, the plurality of apertures 104 are aligned with the plurality of openings 238. In some embodiments, the gasket 224 exerts pressure on the substrate 210 when the substrate holder 201 is in the closed position. The plurality of ribs 218 may be configured to support the substrate 210 and the liner 224. In some embodiments, the pad 224 is positioned in alignment on the surface of the cover 236 by a plurality of ribs. In some embodiments, gasket 224 is positioned such that when substrate 224 rests on top of substrate holder 201 and substrate holder 201 is in the closed position, an airtight seal is formed between gasket 224 and substrate 210. In some embodiments, a hermetic seal is formed between gasket 224 and substrate 210. In some embodiments, the gasket 224 is configured to prevent fluid communication between the plurality of openings 238 when the cover 236 is in the closed position.

The movable tab 262 includes a top portion 268 extending from a substantially vertical wall 270, the wall 270 extending further from the cover 236. The vertical wall 270 engages the body of the non-movable tab 260 (e.g., a mating portion thereof) when the substrate holder 201 is in the closed position. The top portion 268 extends outwardly away from the main body of the movable tab 262 to form a gripping handle for the user. In some embodiments, grasping the handle may assist the user in pulling or pushing the movable tab 262 (e.g., while engaging and/or disengaging the non-movable tab 260). The top portion 268 extends outwardly away from the body of the movable tab 262 at an angle a relative to a horizontal plane parallel to the bottom surface 232 of the cover 236. In some embodiments, the angle α may be about 15 degrees. In some embodiments, the angle α can be at least about 5 degrees to about 90 degrees or more (e.g., between about 5 degrees to 10 degrees, 10 degrees to 15 degrees, 15 degrees to 20 degrees, 20 degrees to 25 degrees, 25 degrees to 30 degrees, 30 degrees to 35 degrees, 35 degrees to 40 degrees, 40 degrees to 45 degrees, 45 degrees to 50 degrees, 50 degrees to 55 degrees, 55 degrees to 60 degrees, 60 degrees to 65 degrees, 65 degrees to 70 degrees, 70 degrees to 75 degrees, 75 degrees to 80 degrees, 80 degrees to 85 degrees, 85 degrees to 90 degrees).

First height h_mIncluding a top portion 268 of the movable tab 262 and a vertical wall 270. Second height h_vIncluding a vertical wall 270 of the movable tab 262. In some embodiments, the first height h of the movable tab 262_mApproximately equal to the height h of the pad 224_g. In some embodiments, the first height h of the movable tab 262_mMay be greater than the height h of the pad 224_gAbout 5% higher. In some embodiments, the first height h of the movable tab 262_mMay be greater than the height h of the pad 224_gAt least about 5% to about 20% (e.g., between about 5% to 10%, 10% to 15%, 15% to 20%). The first height h of the movable tab 262 when the substrate holder 201 is in the closed position_mMay help secure the gasket 224 so that a force is applied to the gasket wall 209 adjacent the locking tab 258.

In some embodiments, the second height h of the movable tab 262_vMay be approximately equal to the height h of the pad 224_g. In some embodiments, the second height h of the movable tab 262_vMay be greater than the height h of the pad 224_gAbout 5% higher. In some embodiments, the second height h of the movable tab 262_vMay be greater than the height h of the pad 224_gAt least about 5% to about 20% (e.g., between about 5% to 10%, 10% to 15%, 15% to 20%) higher. The second height h of the movable tab 262 when the substrate holder 201 is in the closed position_vMay help secure the gasket 224 so that a force is applied to the gasket wall 209 adjacent the locking tab 258.

In the example shown in fig. 2, the movable tabs 206 each include a corresponding opening 281, and the movable tabs 262 include an opening 282. The openings 281 and 282 are vertically extending openings with the non-movable tabs 208, 260 engaged with the movable tab 206 in the closed position of the substrate holder 201. The non-movable tabs 208, 260 project horizontally outward from a side wall of the base 234 (e.g., the first side wall 103 shown in fig. 1). Since the non-movable tabs 208, 260 project horizontally outward, the non-movable tabs 208, 260 contact portions of the movable tabs 206, 262 when the substrate holder 201 is moved from the open position (fig. 2) to the closed position (fig. 1).

For example, when the cover 236 and the base 234 are brought toward each other to move the substrate holder 201 from the open position (fig. 2) to the closed position (fig. 1), the non-movable tab 208 contacts the ramp portion 271 of the movable tab 206 and the non-movable tab 260 contacts the ramp portion 272 of the movable tab 262. This contact deflects the movable tabs 206, 262 outward, moving the non-movable tabs 208, 260 beyond the ramp portions 271, 272 of the movable tabs 206, 262 into the openings 281, 282. In some embodiments, to facilitate deflection of the movable tab 262, a user may operate a grasping handle of the movable tab 262. Once the non-movable tabs 208, 260 are moved beyond the ramp portions 271, 272 into the openings 281, 282, the movable tabs 206, 262 return toward their neutral position such that the surfaces of the movable tabs 206, 262 defining the openings 281, 282 are configured to contact the horizontally protruding portions of the non-movable tabs 206, 262. This creates a locking engagement between the non-movable tabs 208, 260 and the movable tabs 206, 262 that prevents the base 234 and the cover 236 from moving away from each other. When the non-movable tabs 208, 260 are positioned within the openings 281, 282, the substrate holder 201 is in the closed position. In the closed position of the substrate holder, the gasket 224 contacts the substrate (e.g., is compressed against the substrate) and provides a fluid seal between the sample regions of the substrate. In the example shown in fig. 2, the substrate holder 201 includes a plurality of ribs extending perpendicularly or protruding outwardly from the bottom surface 232. The gasket 224 is not in contact with (i.e., does not abut) the plurality of ribs 218. In some embodiments, the liner is not co-molded with the cover. The gasket is mounted to the cover in another suitable manner. In such embodiments, one or more ribs may be adjacent or abutting one or more cushion walls to help hold the cushion in place. In some embodiments, when the cushion is not co-molded with the cover, the one or more ribs may be near or abut 1, 2, 3, or 4 of the cushion walls. In some embodiments, the plurality of ribs 218 delineate a section of the bottom surface 232 (e.g., a section that is sufficiently sized and configured to receive the liner 224). The plurality of ribs 218 may be disposed parallel to the second sidewall 205. In some embodiments, the liner is co-molded with the cover.

The plurality of ribs 218 may have a width w. In some embodiments, as shown in fig. 2, the plurality of ribs 218 may have an equal width w. In some embodiments, the width of the plurality of ribs 218 may vary. One of the ribs 218 extends substantially perpendicularly from the bottom surface 232 near the first end 211a of the substrate holder 201. The remaining ribs 218 extend perpendicularly from the bottom surface 232 near the second end 211b of the substrate holder 201. In some embodiments, the substrate holder 201 can include a plurality of ribs (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, or more ribs). In some embodiments, the substrate holder 201 may include one or more ribs that extend perpendicular to the plurality of ribs 218.

The plurality of ribs 218 may have a height h. In some embodiments, as shown in fig. 2, the plurality of ribs 218 may have equal heights h. In some embodiments, the height h may be provided such that the top edges of the plurality of ribs 218 are in contact with the base 110 (shown in FIG. 1) when the support is in the closed position (FIG. 1). In embodiments where the gasket 224 is compressed against the substrate 110, the contact between the plurality of ribs 218 and the substrate 110 may define a maximum amount of compression of the gasket 224. In particular, when the top edges of the plurality of ribs 218 are in contact with the substrate 110, further compression of the gasket 224 may be prevented. In some embodiments, the plurality of ribs 218 secure the base 110 by contacting a surface of the base 110 when the support is in the closed position (fig. 1). In some embodiments, the first surface 228 of the substrate 110 engages at least one of the plurality of ribs 218 when the substrate holder 101 is in the closed position (fig. 1). In some embodiments, the first surface 228 of the substrate 110 includes a sample region. In some embodiments, the sample region is a section of the substrate configured to receive one or more samples. In some embodiments, the substrate 110 includes one or more sample regions.

Fig. 3 is an exploded view of an exemplary support apparatus 300 in an open position. The cover 336 is configured to receive the pad 324. The cover 336 defines four apertures 342 that receive four feet 344 of the pads 324. Each leg 344 protrudes from a bottom surface 366 of the pad 324. In some embodiments, each leg 344 protrudes from a corner of the gasket 324 (e.g., when two gasket walls 309 meet and form a right angle) or near a corner of the gasket 324. The legs 344 may be composed of a pliable or flexible material (e.g., a polymeric material such as silicone). In some embodiments, the feet 344 have a circular base with a diameter that is larger than the diameter of the holes 342. The legs 344 are configured to partially or fully deform in shape when introduced into the holes 342 such that the legs 344 are secured once inserted and are not easily removed due to the different diameters of the circular base of the legs 344 and the holes 342. In some embodiments, the liner 324 is permanently attached to the cover 336. In some embodiments, the pad 224 can be reversibly attached to the cover 336.

The gasket 324 includes an L-shaped protrusion 326 extending from one of the gasket walls 309. In some embodiments, L-shaped projections 326 extend from the corners of the gasket 324. In some embodiments, the L-shaped protrusion 326 is configured to be received by a groove 340 defined by the cover 336. In some embodiments, groove 340 is an L-shaped groove. In some embodiments, the shape of the recess 340 is complementary to the shape of the L-shaped protrusion 326.

The cover 336 includes a plurality of T-shaped ribs 346 and cross-shaped ribs 348 that extend perpendicularly or project outwardly from the bottom surface 332 of the cover 336. In some embodiments, cover 336 includes eight T-shaped ribs 346. In some embodiments, cover 336 includes at least about one to about eight T-ribs 346 (e.g., 1, 2, 3, 4, 5, 6, 7, or 8T-ribs 346). The T-shaped rib 346 may be spaced from the bottom surface 332 by a length l of the liner 324_gA portion of the alignment extends vertically or projects outwardly. The T-shaped rib 346 may be aligned from the width w of the bottom surface 332 and the liner 324_gA portion of the alignment extends vertically or projects outwardly. In some embodiments, the cover 336 includes at least about one to about three cross-shaped ribs 348 (e.g., 1, 2, or 3 cross-shaped ribs 348). A cross-shaped rib 348 may extend perpendicularly or project outwardly from a portion of the bottom surface 332 where the four apertures 304 meet.

Fig. 4 is a perspective view of a support apparatus 400 comprising a substrate holder 401 and a substrate 410. Substrate holder 401 is shown with cover 436 detached from base 434. The pair of hinges 416 may be breakable hinges. In some embodiments, the pair of hinges 416 may be selectively, manually separable to allow a user to separate the cover 436 from the base 434. In some embodiments, the pair of hinges 419 may include breakable portions. In some embodiments, the breakable portion can comprise a breakable webbing. In some embodiments, the pair of hinges 416 is irreversibly breakable. For example, once broken, the cover 436 and the base 434 cannot be attached via the pair of hinges 416. In some embodiments, the pair of hinges 419 may be reversibly breakable. For example, in some embodiments, the pair of hinges 416 can include a first portion 450 and a second portion 452 that can be configured to reversibly engage one another. In some embodiments, first portion 450 of hinge 416 extends integrally from cover 436, and second portion 452 of hinge 416 extends integrally from base 434. In some embodiments, the first portion 450 and the second portion 452 can directly engage one another via a fastener (e.g., a snap fit, a force fit, or a male-female connection). In some embodiments, there is no hinge 416 on the device.

In some embodiments, one or more sample areas 464 defined on the first surface 428 of the substrate 410 may fall within the enclosed section defined by each of the plurality of openings 438 of the gasket 424. In some embodiments, the substrate 410 can include samples (e.g., biological material samples) on a portion of its surface that are aligned with one or more of the plurality of openings 438. In some embodiments, the sample (e.g., a biological material sample) can be identified in the same manner as the corresponding wells of the plurality of openings 438.

In some embodiments, the base 434 is configured to receive the substrate 410. When the substrate holder 401 is in the closed position, the cover 436 is configured to be placed on top of the base 434 to secure the gasket 424 and the substrate 410 between the cover 436 and the base 434. In some embodiments, the user separates cover 436 from base 434 prior to placing substrate 410 in base 434. Once separated, the user may insert the substrate 410 into the base 434 in the direction of arrow 414. A user may place cover 436, for example, in the direction of arrow 412, on top of base 434 of retention substrate 410 and snap cover 436 onto base 434 via locking tabs 407 and 458 to secure substrate 410.

In some examples, the second surface of the substrate 410 may rest on a portion of the base 434 of the substrate holder 401. For example, base 434 may include lip 422 configured to retain substrate 410 within base 434. For example, the substrate 410 may rest on the lip 422 and remain in place. In some embodiments, the lip 422 extends around the perimeter of the base 434. In some embodiments, the substrate may be secured to the base of the substrate holder. In an example, substrate 410 may be placed in base 434 in the direction of arrow 414. The second surface of the base 410 may be placed in contact with the lip 222 and the first surface 428 of the base 410 in contact with the plurality of ribs 418 and the gasket 424 when the support apparatus 400 is in the closed position. In some embodiments, substrate 410 may be loaded into base 434 without the use of tools. The base 434 may include an opening 420 sized to sufficiently expose one or more portions of the substrate 410. In some embodiments, the opening 420 is sized such that a majority of the second surface of the substrate 410 is exposed and not covered. In some embodiments, the opening 420 may enable the second surface of the substrate 410 to be in contact with a surface of a heating device (e.g., a plate 3110 as described in PCT/US2019/065100, the entire contents of which are incorporated herein by reference). In some embodiments, the first surface 428 of the substrate 410 includes a sample region 464. In some embodiments, base 434 includes an opening 420 that exposes at least a portion of a second surface of substrate 410 when substrate 410 is placed in base 434.

In some embodiments, the movable tabs 406 and 458 may have a C-shaped configuration. In some embodiments, the movable tabs 406 and 458 are constructed of a flexible material that enables them to flex away from the body of the substrate holder to engage the non-movable tabs 408 and 462, respectively. In some embodiments, the non-movable tabs 408 and 462 are not capable of flexing. In some embodiments, the non-movable tabs 408 and 462 are rigid and do not flex when engaged with the movable tabs 406, 458.

In some embodiments, the first sidewall 403 may measure about 3 inches. In some embodiments, the second sidewall 405 may measure about 1 inch. In some embodiments, the substrate 410 may measure approximately 75 millimeters (mm) by 25 mm. In some embodiments, the substrate 410 may measure approximately 75 millimeters (mm) by 50 mm. In some embodiments, the substrate 410 may measure approximately 48 millimeters (mm) by 28 mm. In some embodiments, the substrate 410 may measure approximately 46 millimeters (mm) by 27 mm. In some embodiments, the substrate 410 is a slide.

The support device may be substantially similar in structure and function to the support devices 100, 200, 300, 400 discussed above in several respects, but may include, instead of the plurality of apertures 104, 304 and the plurality of openings 138, 238, 338, 438, an alternative number, size and shape of apertures defined by the cover, and an alternative number, size and shape of cushion openings. In some embodiments, the support device may have two holes. In some embodiments, the support device may have two cushion openings. This difference between the number, size, and shape of the wells and gasket openings can allow a user to use the substrate holder to support a substrate having various numbers, sizes, and shapes of sample regions defined on the surface of the substrate.

FIG. 5 illustrates an example support apparatus 500 in a closed position having two example apertures. The support device 500 includes a first aperture 554a and a second aperture 554 b. The support device 500 includes two gasket openings that align with the first aperture 554a and the second aperture 554b and enable portions of the substrate 510 to be exposed when the substrate holder 501 is in the closed position. In some embodiments, the support device 500 can include a cover 536 defining 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, or more apertures. In some embodiments, the support device 500 includes 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, or more cushion openings. In some embodiments, the support device 500 can include a cover 536 that defines between 1 to 4, 4 to 8, 8 to 16, or 16 to 20 apertures. In some embodiments, the support device 500 includes between 1 to 4, 4 to 8, 8 to 16, or 16 to 20 gasket openings.

In another aspect, the present disclosure includes a method of culturing a sample disposed on a sample region of any of the substrates disclosed herein. In some embodiments, the method comprises mounting the substrate on any of the support devices disclosed herein. The method also includes positioning the substrate and the support device on a heating device (e.g., a laboratory hot plate). The method also includes activating the heating device to transfer heat to the sample (e.g., via a second surface of the substrate exposed via the opening of the substrate holder). In some embodiments, at least 60% of the sample area is covered by the support device when the substrate holder is coupled to the support device. In some embodiments, at least 50%, 40%, 30%, 20%, 10% or less of the sample area of the substrate is covered by the support means.

While the embodiments shown in fig. 1-4 show the non-movable tab (e.g., non-movable tab 108, 160, 208, 260, 408, 462) on the base (e.g., base 134, 234, 334, 434) and the movable tab (e.g., movable tab 106, 162, 206, 262, 406, 460) on the cover (e.g., cover 136, 236, 436), in further embodiments the non-movable tab is on the cover and the movable tab is on the base.

OTHER EMBODIMENTS

It is to be understood that while the utility model has been described in conjunction with the detailed description thereof, the foregoing description is intended to illustrate and not limit the scope of the utility model, which is defined by the scope of the appended claims.

Disclosed are systems, devices (e.g., apparatuses), and methods that can be used for, can be combined with, can be used to prepare, or are products of the disclosed methods. These and other systems, devices, and methods are disclosed herein, and it is understood that combinations, subsets, interactions, groups, etc. of these systems, devices, and methods are disclosed. That is, although specific reference may not have been made to each individual and collective combination and permutation of these systems, apparatus and methods disclosed herein, each is specifically contemplated and described herein. For example, if a particular system, apparatus, or method is disclosed and discussed and a number of systems, apparatuses, or methods are discussed, each and every combination and permutation of these systems, apparatuses, and methods is specifically contemplated unless specifically indicated to the contrary. Likewise, any subset or combination of these is also specifically contemplated and disclosed.

Other aspects, advantages, and modifications are within the scope of the following claims.

Claims (31)

1. A support device, comprising:

a substrate holder, the substrate holder comprising:

a cover comprising opposing first sidewalls, the cover configured to receive a gasket; and

a base comprising opposing first sidewalls, the base configured to receive a sample substrate;

characterized in that said support means further comprise:

a first locking tab and a second locking tab including a movable tab coupled to the first sidewall of one of the cover and the base and a non-movable tab coupled to the first sidewall of the other of the cover and the base, the first locking tab having a length greater than a length of the second locking tab,

wherein the first locking tab extends along a length of the liner, an

Wherein the movable tab is configured to engage with the non-movable tab to releasably secure the cover to the base.

2. The support apparatus of claim 1 wherein the height of the first locking tab is approximately equal to the height of the pad.

3. The support apparatus of claim 1, wherein a length of the first locking tab is approximately equal to a length of the pad.

4. The support device of claim 1, wherein the sample substrate rests on top of the base of the substrate holder.

5. The support apparatus of claim 1, wherein the cover includes a plurality of ribs extending from a surface of the cover, and wherein the pad is positioned in alignment on the surface of the cover by the plurality of ribs.

6. The support device of claim 1, wherein the gasket is positioned such that an airtight seal is formed between the gasket and the sample substrate when the sample substrate is secured by the substrate holder and the substrate holder is in a closed position.

7. The support apparatus of claim 1, wherein the cover includes at least two apertures.

8. The support apparatus of claim 7, wherein the gasket comprises at least two openings, wherein the at least two openings are positioned such that the at least two apertures are aligned with the at least two openings when the sample substrate is secured by the substrate holder and the substrate holder is in a closed position.

9. The support apparatus of claim 8, wherein the gasket is configured to prevent fluid transfer between the at least two openings when the cover is in the closed position.

10. The support device of any one of claims 1-8, wherein the base includes a lip extending around a perimeter of the base, the lip configured to retain the sample substrate within the base.

11. The support device of any one of claims 1-8, wherein the cover comprises a plurality of ribs extending from a surface of the cover, and wherein the sample substrate engages at least one of the plurality of ribs when the substrate holder is in the closed position.

12. The support device of any one of claims 1-8, wherein the base comprises an opening that exposes at least a portion of the sample substrate when the sample substrate is placed in the base.

13. The support apparatus of any of claims 1-8, wherein each of the first sidewalls of the cover is substantially orthogonal to the second sidewalls of the cover.

14. The support device of any one of claims 1-8, wherein the cover is configured to fold onto the base when the substrate holder is in the closed position.

15. The support apparatus of any of claims 1-8, wherein the length of the first locking tab is at least 90% of the length of the cushion.

16. A support device, comprising:

a substrate holder, the substrate holder comprising:

a cover comprising opposing first and second sidewalls, the cover configured to receive a cushion;

a base comprising opposing first and second sidewalls, the base configured to receive a sample substrate, each of the base and the first sidewall of the cover being longer than each of the base and the second sidewall of the cover; and

a locking tab comprising a movable tab and a non-movable tab, one of the movable tab and the non-movable tab coupled to one of the first sidewalls of the cover and the other of the movable tab and the non-movable tab coupled to one of the first sidewalls of the base, the locking tab having a length approximately equal to the length of the gasket, wherein the locking tab extends along the length of the gasket.

17. The support device of claim 16, wherein the sample substrate rests on top of the base of the substrate holder.

18. The support apparatus of claim 16, wherein the cover includes a plurality of ribs extending from a surface of the cover, and wherein the cushion is positionally aligned on the surface of the cover by the plurality of ribs.

19. The support device of claim 16, wherein the gasket is positioned such that an airtight seal is formed between the gasket and the sample substrate when the sample substrate is secured by the substrate holder and the substrate holder is in a closed position.

20. The support apparatus of claim 16 wherein the cover includes an aperture.

21. The support apparatus of claim 16, wherein the cover comprises at least two apertures.

22. The support device of claim 21, wherein the gasket comprises at least two openings, wherein the at least two openings are positioned such that the at least two apertures are aligned with the at least two openings when the sample substrate is secured by the substrate holder and the substrate holder is in a closed position.

23. The support apparatus of claim 22, wherein the gasket is configured to prevent fluid communication between the at least two openings when the substrate holder is in a closed position.

24. The support device of any one of claims 16-23, wherein the sample substrate comprises a glass slide.

25. The support device of any one of claims 16-23, wherein the base includes a lip extending around a perimeter of the base, the lip configured to retain the sample substrate within the base.

26. The support device of any one of claims 16-23, wherein the cover includes a plurality of ribs extending from a surface of the cover, and the sample substrate engages at least one of the plurality of ribs when the substrate holder is in the closed position.

27. The support device of any one of claims 16-23, wherein the base comprises an opening that exposes at least a portion of the sample substrate when the sample substrate is placed in the base.

28. The support apparatus of any one of claims 16-23, wherein each of the first sidewalls of the cover is substantially orthogonal to the second sidewalls of the cover.

29. The support device of any one of claims 16-23, further comprising at least one breakable hinge extending from one of the second side walls of the cover to one of the second side walls of the base such that the cover is configured to fold onto the base to secure the sample substrate and the pad between the cover and the base when the substrate holder is in the closed position.

30. The support device of claim 29, wherein the cover and the base are pivotably connected by at least one hinge extending from one of the second sidewalls of the cover to one of the second sidewalls of the base such that when the substrate holder is in the closed position, the cover is configured to fold over the base to secure the sample substrate and the pad between the cover and the base.

31. The support device of claim 29 wherein the cover and the base are pivotally connected by the at least one breakable hinge.

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