WO2015085175A1

WO2015085175A1 - Cell blocks for cytopathology and surgical pathology specimens

Info

Publication number: WO2015085175A1
Application number: PCT/US2014/068800
Authority: WO
Inventors: Vinod B. Shidham
Original assignee: Shidham Vinod B
Priority date: 2013-12-05
Filing date: 2014-12-05
Publication date: 2015-06-11

Abstract

The disclosure provides kits, devices, and methods to produce cell blocks for cytopathology and surgical pathology specimens. The cell blocks have cells of interest concentrated along one face of the block along with a marker for visual identification of the cells' locations. The kits, devices, and methods facilitate ease, reproducibility, and flexibility in cell block production.

Description

CELL BLOCKS FOR CYTOPATHOLOGY AND SURGICAL

PATHOLOGY SPECIMENS

CROSS REFERENCE TO RELATED APPLICATION

[0001] This application claims priority to United States Provisional Patent Application Serial No. 61/912,389 filed December, 5, 2013, the entire contents of which are incorporated by reference herein.

FIELD OF THE DISCLOSURE

[0002] The disclosure provides kits, devices, and methods to produce cell blocks for cytopathology and surgical pathology specimens along with other specimens such as in research labs and animal sciences including veterinary medicine. The cell blocks have cells of interest concentrated along one face of the cell block along with a marker for visual identification of the cells' locations. The kits, devices, and methods facilitate ease, reproducibility, and flexibility in production.

BACKGROUND OF THE DISCLOSURE

[0003] Histopathology / surgical pathology describes the examination of tissues or tissue sections to diagnose various diseases or conditions. Cytopathology describes the examination of cells or small tissue fragments to diagnose various diseases or conditions. For example, cytopathology can be used to diagnose placental and fetal health, cancers of various organ systems, certain infectious diseases, blood immune cells, and other inflammatory conditions. The most common cytopathology test is the Pap Smear test.

[0004] As the name "Pap Smear" suggests, a number of cytopathological examinations include "smearing" cells onto a slide and examining the smeared cells under a microscope, often following staining of the cells with a diagnostic reagent. While this approach allows numerous types of diagnoses, cell smearing is also associated with diagnostic drawbacks. For example, smears offer a limited number of cells for examination. Additionally, once the cells have been stained with one diagnostic marker, it is difficult to conduct additional diagnostic stains or tests on the same cells at the same time or at later stages in the future. [0005] Cytopathological tests routinely require preparation of cell blocks. In cell blocks, cells for examination are not smeared but instead are embedded within structures, such as paraffin blocks. The structures can then be sliced and the slices are mounted onto glass slides for examination of the embedded cells.

[0006] Cell blocks offer numerous benefits over cytology smears. For example, the cells are not subjected to mechanical disruption that can be associated with smearing. Further, the cell block can be sliced to provide numerous distinct sections of cells and tissue microfragments for analysis. Each slice of a cell or group of cells can be analyzed with different diagnostic methodologies including an ever increasing number of molecular tests. Cell blocks also offer benefits derived from enhanced sampling with additional morphologic features for better interpretation. Additionally, cell blocks can be easily stored / archived and tested at later times with future available tests.

[0007] There are numerous procedures available to make cell blocks; however, most are associated with drawbacks. One drawback of currently available methods is that they can be inappropriate for use with specimens having low cell or tissue fragment numbers. Another drawback comes from uncertainty of where cells for examination may be found within a particular block. While some methods are available that at least partially address these concerns, the approaches are automated and expensive, limiting their flexibility and use by all practitioners. Automated methods also have major disadvantages of fixative-related interference without conformity to most frequently recommended formalin-fixed paraffin embedded cell blocks for immunohistochemistry and/or many continually expanding lists of molecular tests.

SUMMARY OF THE DISCLOSURE

[0008] There is a need for flexible and affordable kits, devices, and methods to prepare high quality cell blocks. The kits, devices, and methods disclosed herein provide flexible and affordable kits, devices, and methods to prepare cell blocks and are appropriate for use with samples that have low cell or tissue numbers. Additionally, the kits, devices, and methods align the cells for examination along a face of a cell block with a marker so that histotechnologists (who cut very thin sections using microtomes from final paraffin embedded cell blocks) can easily understand the location of the cells within the block. This allows for monitoring of the location of the cells such that the technologist does not cut too deep and lose the cells, or does not cut to the level with the cells and completely miss them. It also allows the flexibility of choosing any fixative (commonly recommended is formalin) for any type of protocol.

BRIEF DESCRIPTION OF THE FIGURES

[0009] FIG. 1 provides a depiction of one embodiment of a device disclosed herein.

[0010] FIG. 2 provides a more detailed structure of one embodiment of a mechanism to initiate an exothermic reaction that can be used with the kits, devices, and methods disclosed herein.

[001 1 ] FIG. 3 provides a depiction of one embodiment of a device disclosed herein wherein the device includes a marker release mechanism.

[0012] FIG. 4 provides an overview of one embodiment of a method of using the device depicted in FIG. 1 .

[0013] FIG. 5 provides an overview of one embodiment of a method of using the device depicted in FIG. 3.

DETAILED DESCRIPTION

[0014] Cytopathology describes the examination of cells or small tissue fragments to diagnose various diseases or conditions. Cytopathological examinations include use of cell blocks. In cell blocks, cells for examination are embedded within structures, such as paraffin blocks. The structures can be sliced with each slice providing an opportunity for diagnostic evaluation of the embedded cells.

[0015] Cell blocks offer numerous additional benefits during interpretation of conventional cytopathological cell smears. For example, the cells (such as those from lesions / tumors with fragile cells) are not subjected to mechanical disruption that can be associated with smearing. Further, the cell block can be sliced to provide numerous distinct cell sections for additional analysis. Each slice of a cell or group of cells can be stained / analyzed with different diagnostic reagents. Cell blocks also offer additional benefits derived from histomorphologic input. Additionally, cell blocks can be easily stored / archived and re-tested at later times with future technologies. [0016] Currently available procedures to form cell blocks suffer from drawbacks associated with, (i) inappropriateness for use with specimens having low cell or tissue fragment numbers; (ii) uncertainty of where cells for examination may be found within a particular block; (iii) lack of flexibility of using appropriate fixatives especially with one of the automated options; and/or (iv) cost.

[0017] The present disclosure provides flexible and affordable kits, devices, and methods to prepare high quality cell blocks. The kits, devices, and methods can be low- cost and/or disposable. The kits, devices, and methods achieve alignment of diagnostic material (cells and/or tissue fragments) at a face of the cell block so that a high percentage of cells is located in one area of the block for examination. The kits, devices, and methods also utilize a marker within the cell block so that those examining the cell block can easily locate the level having the highest number of cells within the block.

[0018] In the simplest form, the kits, devices, and methods disclosed herein include a longitudinal reservoir appropriate for use in a centrifuge. The longitudinal reservoir has a substantially flat removable bottom and a mechanism to dislodge the cell block from the bottom of the longitudinal reservoir following formation. In one embodiment, the dislodging mechanism is a plunger-like cap that can be used to push the cell block out of the longitudinal reservoir by positive pressure.

[0019] During formation of the cell block, the cell block substrate is maintained at a desired temperature. The temperature is chosen to maintain the substrate in its sol stage during centrifugation. In particular embodiments, the desired temperature is 40°C - 65°C. In additional embodiments, the desired temperature is 40°C, 41 °C, 42°C, 43°C, 44°C, 45°C, 46°C, 47°C, 48°C, 49°C, 50°C, 51 °C, 52°C, 53°C, 54°C, 55°C, 56°C, 57°C, 58°C, 59°C, 60°C, 61 °C, 62°C, 63°C, 64°C, or 65°C. In another embodiment, the desired temperature is 50°C, 55°C, or room temperature (18°C-22°C). The precise temperature used during a particular cell block formation can depend on the cell block substrate used to form the block. Appropriate cell block substrate media include media available from Biolnnovations, LLC, and various other gels including gelatin, starch, gums, sea weed products, and alginates. [0020] The desired temperature can be achieved and maintained using any appropriate mechanism, for example, a heated centrifuge, microwave, radiant light, water bath, etc.

[0021] In additional embodiments of the kits, devices, and methods disclosed herein, the longitudinal reservoir can be provided within an outer chamber or jacket. In particular embodiments, the outer chamber or jacket can provide thermal insulation for the longitudinal reservoir. In additional embodiments, the outer chamber or jacket can additionally or alternatively house chemical(s) that can be triggered to generate an exothermic reaction. Appropriate chemicals for use include chemical(s) produced by Biolnnovations, LLC. Additional exemplary chemicals include heat retaining liquids such as oils and solutions that reach a particular temperature by microwave or conduction heating.

[0022] In a particular embodiment, the one or more chemicals are contained within an airtight pouch or compartment. In particular embodiments, the one or more chemicals can be contained in a sealed pouch or within an air tight compartment of the outer chamber or jacket.

[0023] The exothermic reaction can be triggered upon exposure to sudden energy release, upon mechanical agitation, and/or upon mechanical shearing, leading to a temperature change. The exothermic reaction can begin when the pouch or compartment releases reaction initializing crystals within it. Release of crystals can be based on the engagement of a knob or plunger that dislodges, disforms, or pierces a portion of the pouch or compartment. In one embodiment, the exothermic reaction is initiated by tightening a screw-like device referred to herein as a heat activation turning knob. In this embodiment, tightening the knob dislodges a seal of the pouch or compartment allowing the entry of crystals to initiate the exothermic reaction. Accordingly, the outer chamber or jacket can assist with achieving and/or maintaining a desired temperature of the cell block during its formation. An outer chamber is rigid in form whereas a jacket can be formed from a flexible, malleable, or expandable material.

[0024] FIG. 1 provides a depiction of an exemplary device disclosed herein. In this embodiment, there is a longitudinal reservoir 1 with a substantially flat removable bottom; a cell block dislodging mechanism in the form of a plunger-like cap 2; an outer chamber 3 containing chemicals 4 to generate an exothermic reaction; a mechanism to generate an exothermic reaction in the form of a heat activation turning knob 5; and a removable base 6 allowing exposure and removal of the cell block while retaining residual chemicals in the outer chamber. The top panel of FIG. 1 depicts the device before use of the heat activation turning knob 5 whereas the bottom panel of FIG. 1 depicts the device after use of the heat activation turning knob 5. As is understood by one of ordinary skill in the art, in this embodiment, the removable base provides the substantially flat bottom for the longitudinal reservoir. In alternative embodiments, the removable base and substantially flat bottom can be different separately removable pieces of the device.

[0025] FIG. 2 provides a more detailed structure of a mechanism to generate an exothermic reaction in the form of a heat activation turning knob that can be used with the kits, devices, and methods disclosed herein. In the depicted embodiment, turning the knob 20 tightens down a screw 30 that dislodges, deforms, or pierces a covering 40 leading to release of crystals for initiation of the exothermic reaction. In the depicted embodiment, the covering can be a material such as a plastic film or bag housing crystals 50. Puncturing the material releases the crystals into the outer chamber thereby initiating an exothermic reaction.

[0026] Kits, devices, and methods disclosed herein can also optionally include a marker release mechanism. FIG. 3 depicts an exemplary embodiment with a marker release mechanism 20. In embodiments adopting a marker release mechanism, the marker can optionally be provided pre-loaded into the release mechanism. Alternatively, a user of the kits, devices, and methods can place a marker into the marker release mechanism during use. At the appropriate time during the disclosed methods (described in more detail below), the release mechanism can be activated as shown in FIG. 3. Particularly, the inverted jar like structure pierces through a membrane and releases the marker, which was stored in the chamber prior to the piercing of the membrane, by the pushing down edge of the inverted jar which also act as a storing chamber for the marker.

[0027] The marker described herein can include any device that is visually detectable within a cell block. Generally, the marker can be a dark-colored opaque beacon-like marker which facilitates cutting the cell block by allowing the monitoring of the depth of cutting into the area with maximum number of cells-tissue fragments at the level when the marker is just exposed while cutting the block. Particularly useful markers include those that are formed from protein and/or cellulose material with dark colors.

[0028] The described kits, devices, and methods achieve relatively cellular cell block sections from relatively hypocellular specimens with singly scattered or small groups of cells or a few microfragments of tissue. The described cell block sections can be achieved by concentrating diagnostic material along a cutting face of cell block using a centrifugation step wherein the gel is kept molten during centrifugation by a heating mechanism. In particular embodiments, the heating mechanism is an exothermic reaction in the outer chamber or jacket of the device. The section cutting is further enhanced to get good cellular cell block sections guided by appropriate depth of cutting using a marker, for example an Anjani-Vinod (AV) marker. See Varsegi & Shidham (2009). Cell Block Preparation from Cytology Specimen with Predominance of Individually Scattered Cells. J Vis Exp. (JoVE- Journal of Visualized Experiments) 2009 Jul 21 ;(29). pii: 1316. JoVE. 29. doi: 10.3791/1316. PMID: 19623160.

[0029] One exemplary method is described in FIG. 4 which depicts use of an embodiment depicted in FIG. 1 . FIG. 4 provides associated narrative of the depicted method. An additional exemplary method is described in FIG. 5 which depicts use of an embodiment depicted in FIG. 3. FIG. 4 provides associated narrative of the depicted method.

[0030] Particular method embodiments include an initial preparation step to concentrate a specimen for analysis. In this embodiment, a concentrated button of sediments in the specimen is created by centrifuging the specimen at an appropriate G sufficient to sediment diagnostic material completely (1000 to 2000 rpm) in reasonable time (2 to 5 minutes). This is followed by discarding the supernatant (for example, by inverting the centrifugation tube in discard container). Note that if the specimen under consideration is fatty tissue with density lower than water, this tissue, such as fibroadipose tissue fragments of anterior fat pad aspirate, cannot be centrifuged to concentrate diagnostic fibroadipose tissue fragments (instead fibrofatty specimens need to be filtered and processed by a different protocol).

[0031] Next the sediment can be suspended in the residual supernatant retained by capillary action along the tube wall, which accumulates after keeping the tube upright for a while. This concentrated specimen can be used for preparing a cell block as disclosed herein.

[0032] A solidified aliquot of gel can be made molten by heating it. In particular embodiments, the heating can be at 100^°C in a water bath or microwave (properly timed under proper precautions). Before use, the molten gel can be cooled to 55^°C.

[0033] Next, concentrated sediment suspension with cells and/or microfragments of tissue can be added to a longitudinal reservoir. Markers can also be added to the longitudinal reservoir.

[0034] The sample for analysis, marker, and longitudinal reservoir can be raised to, and maintained at, a temperature of 55^°C. Cell block substrate / medium cooled to 55^°C can next be added to the sample and marker combination. The longitudinal reservoir and its contents can then be centrifuged at an appropriate G sufficient to sediment diagnostic material completely (e.g., 1000 to 2000 rpm) in a reasonable time (e.g., 2 to 5 minutes).

[0035] After centrifugation, the substrate / medium should be allowed to solidify without disturbing the sedimented layer at the bottom. In a standard refrigerator, solidification generally takes 15-30 minutes. For the best outcome, the substrate / cell block should not be frozen.

[0036] Once solidified, the bottom of the longitudinal reservoir can be removed leaving a cell block with cells of interest along the bottom of the block. If the unscrewing itself does not pull the solidified gel out with it, then it can be pushed out using a cell block dislodging mechanism. In particular embodiments, this is accomplished by pressing a plunger cap after closing the hole in the center of the plunger-like cap handle, with, for example, a fingertip or other convenient object. The solidified gel button should be placed into a container with appropriate fixative or saline.

[0037] If the solidified gel button is taller (thicker) than 2.5 mm, it can be trimmed to 2.5 mm from the bottom up (the bottom has the marker and the cells of interest). The trimming may be facilitated by a trimming device provided with the kits disclosed herein. The cell block can be wrapped in tissue processing paper; placed in a labeled tissue cassette; immersed in a fixative of choice (usually 10% formalin); and processed on a tissue processor after fixation for 2 hours.

[0038] The processed and fixed cell block in the cassette can be paraffin- embedded. The paraffin-embedded cell block can be cut as usual. The marker can guide cutting of the final paraffin block by monitoring the depth of cutting. The area with a maximum number of cells-tissue fragments can be mostly at the level when the marker is just exposed while cutting the block. If serial sections are to be followed in sequential fashion, the marker can further assist as a reference point while evaluating the sections by a Subtractive Coordinate Immunoreactivity Pattern (SCIP) immunohistochemistry approach. See, for example, Shidham & Atkinson; Editors 'Cytopathologic Diagnosis of Serous Fluids' First edition, Elsevier (W. B. Saunders Company); 2007. Ch 5, pp. 55-78.

[0039] The kits disclosed herein can include one or more of the described components allowing the easy formation of high quality cell blocks. For example, the kits can include one or more of (1 ) a longitudinal reservoir with a substantially flat removable bottom; (2) a cell block substrate / medium; (3) a marker; (4) chemical(s) to generate an exothermic reaction; (5) an outer chamber or jacket; (6) a cell block dislodging mechanism; (7) a mechanism to initiate an exothermic reaction; (8) a marker release mechanism; (9) gel button trimming device; and/or (10) diagnostic cell staining reagent(s).

[0040] In various embodiments, the kits described herein further include instructions for using the kit in the methods disclosed herein. The kit may include instructions regarding preparation of cell block substrate / medium, markers, etc.; appropriate reference levels to interpret results associated with using the kit; proper disposal of the related waste; and the like. The instructions can be in the form of printed instructions provided within the kit or the instructions can be printed on a portion of the kit itself. Instructions may be in the form of a sheet, pamphlet, brochure, CD-Rom, or computer- readable device, or can provide directions to instructions at a remote location, such as a website. The instructions may be in English and/or in any national or regional language. [0041] In various embodiments, the packaging, components, and/or instructions are combined into a small, compact kit with printed instructions for use. In various embodiments in which more than one diagnostic cell staining reagents and/or cell block substrates is provided, suggestions for use of the diagnostic cell staining reagents and/or cell block substrates can be labeled in the kit.

[0042] In various embodiments, the kits described herein include some or all of the necessary supplies needed to use the kit effectively, thereby eliminating the need to locate and gather such supplies. Such supplies can include tubes, tube racks, sterilizing fluid, facemask, cytopathology slides, pipette tips, pipettes, gloves, and the like. Variations in contents of any of the kits described herein can be made.

[0043] The Examples below are included to demonstrate particular embodiments of the disclosure. Those of ordinary skill in the art should recognize in light of the present disclosure that many changes can be made to the specific embodiments disclosed herein and still obtain a like or similar result without departing from the spirit and scope of the disclosure.

EXEMPLARY EMBODIMENTS

1 . A device including (i) a longitudinal reservoir with a substantially flat removable bottom and (ii) a cell block dislodging mechanism.

2. A device according to embodiment 1 wherein the cell block dislodging mechanism is a plunger.

3. A device according to any one of embodiments 1 or 2 wherein the longitudinal reservoir is housed within an outer chamber or jacket.

4. A device according to any one of embodiments 1 -3 wherein the outer chamber or jacket houses chemicals to initiate an exothermic reaction.

5. A device according to any one of embodiments 3 or 4 wherein the outer chamber or jacket includes an opening for engagement of an exothermic reaction initiating mechanism.

6. A device according to embodiment 5 wherein the exothermic reaction initiating mechanism is a screw. A device according to any one of embodiments 1 -6 further including a marker release mechanism.

A kit for the non-automated or automated production of cell blocks including (i) a longitudinal reservoir with a substantially flat removable bottom; (ii) a cell block dislodging mechanism; (iii) a cell block substrate; and (iv) a marker.

A kit according to embodiment 8 further including a gel button trimming device. A kit according to any one of embodiments 8 or 9 further including an outer chamber or jacket.

A kit according to embodiment 10 wherein the outer chamber or jacket houses chemicals to initiate an exothermic reaction.

A kit according to any one of embodiments 10 or 1 1 wherein the outer chamber or jacket includes an opening for engagement of an exothermic reaction initiating mechanism.

A kit according to embodiment 12 wherein the exothermic reaction initiating mechanism is a screw.

A kit according to any one of embodiments 8-13 further including a marker release mechanism.

A kit according to any one of embodiments 8-14 further including instructions to prepare a cell block using components of the kit.

A kit according to any one of embodiments 8-15 further including one or more diagnostic cell staining reagents.

A kit according to any one of embodiments 8-16 further including one or more of cytopathology slides or pipette tips.

A method of forming a cell block including utilizing the devices or kits of any one of embodiments 1 -17.

A method of forming a cell block with diagnostic material and a marker concentrated along one face of the cell block including:

loading diagnostic material for analysis, a marker and a cell block substrate into a longitudinal reservoir with a substantially flat removable bottom;

maintaining the longitudinal reservoir and its loaded contents within a temperature range that maintains the cell block substrate in its sol phase; centrifuging the longitudinal reservoir and its loaded contents during the temperature range maintenance step; and

allowing the longitudinal reservoir and its loaded contents to cool below the temperature range thereby forming the cell block.

20. A method according to embodiment 19 further including removing the substantially flat removable bottom of the longitudinal reservoir.

21 . A method according to any one of embodiments 19 or 20 further including removing the formed cell block from the longitudinal reservoir.

22. A method according to embodiment 21 wherein the removing occurs through plunging a plunger to expel the cell block from the bottom of the longitudinal reservoir.

23. A method according to any one of embodiments 19-22 wherein the temperature range is 45°C - 60°C.

24. A method according to any one of embodiments 19-22 wherein the temperature range is room temperature to 48°C to 57°C.

25. A method according to any one of embodiments 19-22 wherein the maintenance of the temperature range occurs by initiating an exothermic reaction in an outer chamber or jacket surrounding a longitudinal reservoir and its loaded contents.

26. A method according to embodiment 25 wherein the exothermic reaction initiation occurs through the turning of a knob associated with a screw.

[0044] As will be understood by one of ordinary skill in the art, each embodiment disclosed herein can comprise, consist essentially of, or consist of its particular stated element, step, ingredient or component. Thus, the terms "include" or "including" should be interpreted to recite: "comprise, consist of, or consist essentially of." As used herein, the transition term "comprise" or "comprises" means includes, but is not limited to, and allows for the inclusion of unspecified elements, steps, ingredients, or components, even in major amounts. The transitional phrase "consisting of excludes any element, step, ingredient or component not specified. The transition phrase "consisting essentially of limits the scope of the embodiment to the specified elements, steps, ingredients or components and to those that do not materially affect the embodiment. As used herein, a material effect would prevent the non-automated formation of a cell block with cells for analysis and a marker concentrated along one face of the cell block.

[0001] Unless otherwise indicated, all numbers used in the specification and claims are to be understood as being modified in all instances by the term "about." Accordingly, unless indicated to the contrary, the numerical parameters set forth in the specification and attached claims are approximations that may vary depending upon the desired properties sought to be obtained by the present invention. At the very least, and not as an attempt to limit the application of the doctrine of equivalents to the scope of the claims, each numerical parameter should at least be construed in light of the number of reported significant digits and by applying ordinary rounding techniques. When further clarity is required, the term "about" has the meaning reasonably ascribed to it by a person skilled in the art when used in conjunction with a stated numerical value or range, i.e. denoting somewhat more or somewhat less than the stated value or range, to within a range of ±20% of the stated value; ±19% of the stated value; ±18% of the stated value; ±17% of the stated value; ±16% of the stated value; ±15% of the stated value; ±14% of the stated value; ±13% of the stated value; ±12% of the stated value; ±1 1 % of the stated value; ±10% of the stated value; ±9% of the stated value; ±8% of the stated value; ±7% of the stated value; ±6% of the stated value; ±5% of the stated value; ±4% of the stated value; ±3% of the stated value; ±2% of the stated value; or ±1 % of the stated value.

[0002] Notwithstanding that the numerical ranges and parameters setting forth the broad scope of the invention are approximations, the numerical values set forth in the specific examples are reported as precisely as possible. Any numerical value, however, inherently contains certain errors necessarily resulting from the standard deviation found in their respective testing measurements.

[0003] The terms "a," "an," "the" and similar referents used in the context of describing the invention (especially in the context of the following claims) are to be construed to cover both the singular and the plural, unless otherwise indicated herein or clearly contradicted by context. Recitation of ranges of values herein is merely intended to serve as a shorthand method of referring individually to each separate value falling within the range. Unless otherwise indicated herein, each individual value is incorporated into the specification as if it were individually recited herein. All methods described herein can be performed in any suitable order unless otherwise indicated herein or otherwise clearly contradicted by context. The use of any and all examples, or exemplary language (e.g., "such as") provided herein is intended merely to better illuminate the invention and does not pose a limitation on the scope of the invention otherwise claimed. No language in the specification should be construed as indicating any non-claimed element essential to the practice of the invention.

[0004] Groupings of alternative elements or embodiments of the invention disclosed herein are not to be construed as limitations. Each group member may be referred to and claimed individually or in any combination with other members of the group or other elements found herein. It is anticipated that one or more members of a group may be included in, or deleted from, a group for reasons of convenience and/or patentability. When any such inclusion or deletion occurs, the specification is deemed to contain the group as modified thus fulfilling the written description of all Markush groups used in the appended claims.

[0005] Certain embodiments of this invention are described herein, including the best mode known to the inventors for carrying out the invention. Of course, variations on these described embodiments will become apparent to those of ordinary skill in the art upon reading the foregoing description. The inventor expects skilled artisans to employ such variations as appropriate, and the inventors intend for the invention to be practiced otherwise than specifically described herein. Accordingly, this invention includes all modifications and equivalents of the subject matter recited in the claims appended hereto as permitted by applicable law. Moreover, any combination of the above- described elements in all possible variations thereof is encompassed by the invention unless otherwise indicated herein or otherwise clearly contradicted by context.

[0006] Furthermore, numerous references have been made to publications, patents and/or patent applications (collectively "references") throughout this specification. Each of the cited references is individually incorporated herein by reference for their particular cited teachings.

[0007] The particulars shown herein are by way of example and for purposes of illustrative discussion of the preferred embodiments of the present invention only and are presented in the cause of providing what is believed to be the most useful and readily understood description of the principles and conceptual aspects of various embodiments of the invention. In this regard, no attempt is made to show structural details of the invention in more detail than is necessary for the fundamental understanding of the invention, the description taken with the drawings and/or examples making apparent to those skilled in the art how the several forms of the invention may be embodied in practice.

[0008] Definitions and explanations used in the present disclosure are meant and intended to be controlling in any future construction unless clearly and unambiguously modified in the examples or when application of the meaning renders any construction meaningless or essentially meaningless. In cases where the construction of the term would render it meaningless or essentially meaningless, the definition should be taken from Webster's Dictionary, 3rd Edition or a dictionary known to those of ordinary skill in the art, such as the Oxford Dictionary of Biochemistry and Molecular Biology (Ed. Anthony Smith, Oxford University Press, Oxford, 2004).

[0009] In closing, it is to be understood that the embodiments of the invention disclosed herein are illustrative of the principles of the present invention. Other modifications that may be employed are within the scope of the invention. Thus, by way of example, but not of limitation, alternative configurations of the present invention may be utilized in accordance with the teachings herein. Accordingly, the present invention is not limited to that precisely as shown and described.

Claims

CLAIMS What is claimed is:

1 . A kit for the non-automated or automated production of cell blocks comprising (i) a longitudinal reservoir with a substantially flat removable bottom; (ii) a cell block dislodging mechanism; (iii) a cell block substrate; and (iv) a marker.

2. A kit according to claim 1 further comprising a gel button trimming device.

3. A kit according to claim 1 further comprising an outer chamber or jacket.

4. A kit according to claim 3 wherein the outer chamber or jacket houses chemicals to initiate an exothermic reaction.

5. A kit according to claim 3 wherein the outer chamber or jacket comprises an opening for engagement of an exothermic reaction initiating mechanism.

6. A kit according to claim 5 wherein the exothermic reaction initiating mechanism is a screw.

7. A kit according to claim 1 further comprising a marker release mechanism.

8. A kit according to claim 1 further comprising instructions to prepare a cell block using components of the kit.

9. A kit according to claim 1 further comprising one or more diagnostic cell staining reagents.

10. A kit according to claim 1 further comprising one or more of cytopathology slides or pipette tips.

1 1 .A device comprising (i) a longitudinal reservoir with a substantially flat removable bottom and (ii) a cell block dislodging mechanism.

12. A device according to claim 1 1 wherein the cell block dislodging mechanism is a plunger.

13. A device according to claim 1 1 wherein the longitudinal reservoir is housed within an outer chamber or jacket.

14. A device according to claim 13 wherein the outer chamber or jacket houses chemicals to initiate an exothermic reaction.

15. A device according to claim 13 wherein the outer chamber or jacket comprises an opening for engagement of an exothermic reaction initiating mechanism.

16. A device according to claim 15 wherein the exothermic reaction initiating mechanism is a screw.

17. A device according to claim 1 1 further comprising a marker release mechanism.

18. A method of forming a cell block comprising utilizing the devices or kits of any one of claims 1 -17.

19. A method of forming a cell block with diagnostic material and a marker concentrated along one face of the cell block comprising:

maintaining the longitudinal reservoir and its loaded contents within a temperature range that maintains the cell block substrate in its sol phase;

centrifuging the longitudinal reservoir and its loaded contents during the temperature range maintenance step; and

20. A method according to claim 19 further comprising removing the substantially flat removable bottom of the longitudinal reservoir.

21 . A method according to claim 19 further comprising removing the formed cell block from the longitudinal reservoir.

22. A method according to claim 21 wherein the removing occurs through plunging a plunger to expel the cell block from the bottom of the longitudinal reservoir.

23. A method according to claim 19 wherein the temperature range is 45°C - 60°C.

24. A method according to claim 19 wherein the temperature range is room temperature to 48°C to 57°C.

25. A method according to claim 19 wherein the maintenance of the temperature range occurs by initiating an exothermic reaction in an outer chamber or jacket surrounding a longitudinal reservoir and its loaded contents.

26. A method according to claim 25 wherein the exothermic reaction initiation occurs through the turning of a knob associated with a screw.