Cell pathology tubes and associated cell processing methods

Cell tubes that can be used both for pathology collection and subsequent cell processing include a tube with a cell bed at a lower portion of the tube. The tube can include a base member that can be detachable from the tube body. The tubes can be used to form cell (cytology) blocks that incorporate the cell bed. The cell bed can be an inert cell bed of paraffin.

FIELD OF THE INVENTION

This invention relates to cell pathology tubes for cell collection and/or processing.

BACKGROUND OF THE INVENTION

Compared to core or open biopsies, fine needle aspirates (FNA) are a quick and relatively safe method of biopsy to provide samples for evaluation of clinically suspicious mass lesions. FNA are often a first step in evaluating whether a patient has a malignancy. Routine collections during FNA include paired smears (Diff-Quick and H&E) and cell block (clot block or cell disk). Each collection is from a single pass or aspiration. The paired smears are routinely performed on most of the passes to rapidly evaluate the morphology of the suspicious lesion. The cell block is used to process the specimen in a manner to allow for ancillary testing. For example, the cell block can be used to evaluate immunohistochemistry, FISH, PCR and the like, as well as to assess morphology. The morphology of blocks may be diagnostically complimentary in that they often provide more data with regard to the architectural arrangement of cells compared to smears. The evaluation of cells from FNAs typically employs very limited cell material. It is a known problem that the quality of the cell block can deteriorate compared to paired cell smears from the same procedure. Unfortunately, this deterioration can impair additional, sometimes confirmatory, testing or analysis that can be carried out on the specimen. This can result in an inconclusive diagnosis typically requiring additional diagnostic procedures at increased risk and/or cost to the patient and may delay specific therapy.

In the past, the cell blocks have been prepared using samples that are expelled onto glass slides and placed in formalin within a short time frame. The cell block is allowed to “clot” together to make a relatively solid specimen that can be processed to make formalin-fixed paraffin embedded (FFPE) tissue samples. This cell block preparation process can result in undue cell loss resulting in decreasing (and sometimes no) cell yields in the FFPE tissue samples.

SUMMARY OF EMBODIMENTS OF THE INVENTION

Embodiments of the invention provide tubular devices with integrated cell beds (e.g., paraffin beds) that can directly receive cells from a FNA or other sources at a collection site without requiring the use of a glass slide.

Embodiments of the invention provide tubular devices that can be used for the collection, transfer and subsequent centrifuge processing of tissue samples to form a cell block with suspended cells.

Aspects of the invention are directed to methods of collecting and processing a biosample. The methods include: (a) providing a tubular container with an internal cavity having a cell bed residing above a closed bottom end of the container; (b) inserting a biosample comprising cells into the tubular container so that cells reside on the cell bed; (c) placing the tubular container with the cell bed and biosample in a centrifuge; (d) centrifuging the biosample in the tubular container so that cells from the biosample deposit as a pellet against the cell bed; (e) inserting a liquid matrix material in the tubular container above the cell bed; (f) forming a solid cell block of the liquid matrix material holding distributed cells therein above the cell bed; and (g) removing the solid cell block with the cell bed from the tubular container.

The cell bed can be a solid, shape-changeable, moldable material that is able to change in shape in response to forces above those applied during the centrifuging step and can retain that shape during the inserting, centrifuging, forming and removing steps.

The cell bed can include solid paraffin and extends across the tubular container.

The cell bed can have a middle portion with a substantially conical shape that merges into an outer cylindrical upwardly extending outer portion that conformably attaches to an inner surface of a sidewall of the tube and the cell bed defines a closed solid cell bed surface.

Inserting the biosample can be carried out by depositing cells from a fine needle aspirate directly from a needle holding the aspirate onto the cell bed.

Placing the tubular container in the centrifuge can optionally be carried out by first placing the tubular container holding the biosample on the cell bed in a coupler, adapter or larger tube forming a centrifuge assembly, then placing the assembly in a bucket of the centrifuge and centrifuging the assembly.

The cell block can include cells from a fine needle aspirate tissue sample. The method can include, before the centrifuging step, inverting the tubular container with the cell bed, then attaching the tubular container to an open end of an elongate body comprising a liquid, then inverting the tubular container with the cell bed while attached to the elongate body with the liquid so that the cell bed is at a lower end of the tubular container, then centrifuging the biosample in the tubular container.

Other embodiments are directed to cell pathology containers. The containers can have a tubular body having an open interior space and open opposing first and second end portions. A base is removably attached to the second end portion of the tubular body. A solid cell bed resides in the tubular body proximate the base.

The cell bed can have a substantially planar bottom.

The cell bed can have a solid, shape-changeable, moldable material that is able to retain a defined conical or frustoconical shape.

The cell bed can include solid paraffin and extend across the tubular container to define a closed cell bed surface.

The cell bed can have a middle portion with a substantially conical shape.

The cell bed can have a middle portion that merges into an outer cylindrical upwardly extending outer portion that conformably attaches to an inner surface of a sidewall of the tube and the cell bed can define a closed solid cell bed surface.

The solid cell bed can be pre-formed in the tubular body and/or base and can define a closed solid cell bed surface, and wherein the container is enclosed in sterile packaging.

The base can include an internal substantially planar surface that holds the cell bed.

The base can hold a spacer that rises above a lower portion of the base and extends into the tubular body, and the spacer can include an upper substantially planar surface that holds the cell bed.

The base can include an annular recess that surrounds the planar surface and engages a lower portion of the tubular body.

The tubular body first portion can releasably attach to either or both of a cap or elongate body having a length that is greater than that of the tubular body.

The base and cap can have respective ledges of substantially common diameter that extend radially outward from a centerline of the container to reside a distance beyond a diameter of the tubular body.

The container can have a volume that is between about 10 mL to about 100 mL. The tubular body has threads on upper and lower portions thereof, the lower portion configured to threadably attach to the base.

The tubular body can be sterile and configured to hold human or animal cell samples.

The first end portion of the tubular body can be attached to an open end of the elongate body. The elongate body can have a tapered segment that merges into a lower segment having a greater outer diameter. The elongate body can have a removable end cap on an end opposing the open end.

Embodiments of the invention provide cell disks that can be used to make pathology or diagnostic specimens, e.g., surgical pathology FFPE specimens from any kind of cellular source, e.g., suspension. This applies to body fluids (pleural, pericardial, lung washing, etc.). These fluids have diagnostic utility, but also might be the only specimen in certain cases.

Special stains for amyloid or microorganisms can also be performed on any collected specimen, e.g., any FFPE specimens. This can be adapted to fluids for diagnosis.

Other aspects are directed to methods of collecting a biosample. The methods include: (a) providing a tubular container with an internal cavity having a pre-formed solid paraffin cell bed residing above a closed bottom end of the container; (b) inserting a needle with a fine needle aspirate sample comprising cells into the tubular container so that cells reside on the cell bed; and either (c)(i) placing a cap on the container before or after inserting the biosample or (c)(ii) attaching the tubular container with the cell bed on an upper portion of an elongate body with liquid and cells from other passes of FNA.

Yet other methods are directed to methods of processing a biosample. The methods include: (a) obtaining a tubular container with an internal cavity having a solid cell bed residing above a closed bottom end of the container and a biosample comprising cells that reside on the cell bed, wherein the tubular container with the solid cell bed and biosample is obtained from a collection site; (b) centrifuging the biosample in the obtained tubular container so that cells from the biosample deposit as a pellet against the cell bed; (c) inserting a liquid matrix material in the tubular container above the cell bed; (d) forming a solid cell block of the liquid matrix material holding distributed cells therein above the cell bed; and (e) removing the solid cell block with the cell bed from the tubular container.

Still other aspects of the invention are directed to methods of making cell beds for cell pathology containers. The methods include forming solid paraffin into a defined cell bed shape so that the cell bed has a middle portion with a substantially conical or frustoconical shape that merges into an outer cylindrical upwardly extending outer portion that conformably attaches to an inner surface of a sidewall of the tube and the cell bed defines a closed solid cell bed surface.

The tubular container holding the biosample on the cell bed can include an elongate body attached to an upper end thereof. The elongate body can include liquid with supplemental cells from needles used to obtain additional fine needle aspirates of target tissue corresponding to the biosample on the cell bed. The centrifuging can be carried out to form a pellet of the biosample cells and the supplemental cells.

DETAILED DESCRIPTION OF EMBODIMENTS OF THE INVENTION

Like numbers refer to like elements throughout. In the figures, the thickness of certain lines, layers, components, elements or features may be exaggerated for clarity. Broken lines illustrate optional features or operations unless specified otherwise. One or more features shown and discussed with respect to one embodiment may be included in another embodiment even if not explicitly described or shown with another embodiment.

The term “about” means that the recited number or value can vary by +/−20%.

The term “biosample” refers to human or animal tissue, blood or other solid or liquid samples that have cellular material. The cellular material can be limited cellular material, obtained from an FNA or other specimens including, for example, washings, lavages, and endoscopic procedures. Embodiments of the invention can be used for immunohistochemistry (IHC) or other studies including RNA and DNA analysis, research or studies including FISH, PCR and the like and/or to assess morphology. Embodiments of the invention may be used with or for stains, such as “special stains” like Gram stains, Reticulin, Mucin and may others as is well known to those of skill in the art.

Embodiments of the invention provide cell disks that can be used to make surgical pathology FFPE specimens from any kind of suspension. This applies to body fluids (pleural, pericardial, lung washing, etc.). These fluids have diagnostic utility, but also might be the only specimen in certain cases.

Special stains for amyloid or microorganisms can also be performed on any collected specimen, e.g., any FFPE specimens. This can be adapted to fluids for diagnosis.

Embodiments of the invention may be suitable for veterinarian use, medical human use or research for human and/or with laboratory animals.

The term “sterile” means that the noted device or material meets or exceeds defined medical guidelines of cleanliness and is substantially (if not totally) without contaminants so as to be suitable for medical uses (e.g., diagnosis).

Turning now to the figures,FIG. 1Aillustrates a cell collection tube10with a base11, an internal cavity10cand an internal cell bed20held proximate the bottom10lof the tube10. The cell bed10can be an inert cell bed. The term “inert cell bed” refers to a solid material that can support processed cell material above the base11, typically in a block form, while preserving the cells, typically without chemically interacting with the cells. Post collection and after processing, the cell bed20with patient cells C can be removed, substantially intact, e.g., scraped, pushed or otherwise disengaged from the base11without disrupting the collected cells thereon for cell evaluation.

As shown inFIG. 1A, the cell bed20can be substantially planar (e.g., the top and bottom can be flat similar to a flat block). As shown inFIG. 1B, the cell bed20can have an upper portion20uwith a defined three-dimensional body shape20s. The body shape20scan include a substantially conical or frustoconical center portion20cfor facilitating cell collection during centrifugation. The center portion20ccan taper to have a valley that holds the cells/pellet100. The cell bed20can extend across an entire interior cavity space10cat the bottom portion of the tube10lto define a closed surface cell bed above the base11. In some embodiments, the cell bed20can include an upwardly extending outer sidewall20wthat rises a distance above the laterally extending portion and conforms to the inner surface10iof the tube sidewall10wat the bottom portion of the tube10lThe outer perimeter sidewalls20wcan inhibit some samples from being attracted to hydrophilic plastic (polymer) walls of the cylinder10bthat can facilitate forming the cell plug in the desired cell disk or block form. In addition, or alternatively, the taller walls20wcan stay with the cylinder body10bwhen separated from the base11, with the lower portion of the cell bed20intact and remaining attached to the walls20w. In other embodiments, the cell bed walls20wmay detach from the lower cell bed20when the base11is removed.

In some embodiments, the tube body10b, base11and cap12can comprise a molded polymeric material that is sterilized for use. The tube body10b, base11and/or cap12may also comprise other suitable materials, including, for example, glass.

In some embodiments, the cell bed20has a lower surface20sthat can be substantially planar20p. The base11can also have an internal surface11ithat is substantially planar. However, the base surface11ican have other shapes and may include narrow channels or slots or can include “waves” or dimples and the like. The surface11ican be configured to allow for ease of removal of the cell bed20with cells in a “cell block” form100(FIGS. 4A, 4B), post-processing. The cell bed lower surface20scan reside against the base surface11ior may reside a distance above the base11attached to an inner surface10sof a sidewall10wof the tube10.

The internal base surface11ican include a non-stick material and/or coating that reduces sliding friction and/or otherwise facilitates the removal of the cell bed20from the base11with the cell block100(FIG. 4C) for conventional cell evaluation after processing. In some embodiments, the cell bed20with the cell block100can be pushed off the base or pushed up and out of the tube body once the base is removed. Optionally, a thin flexible liner110(FIG. 4C) can reside on the surface11ito allow the cell block100to be lifted or more readily slid off the base11(FIG. 4C). The liner110, where used, can be adhesively attached to the internal surface of the base11iand a user can lift to peel an edge and dislodge the cell block100.

The cell bed20can be a monolithic solid layer of an inert material. In other embodiments, the cell bed20can comprise a plurality of stacked layers of different solid materials or a mixture of materials. The cell bed20can comprise paraffin or other suitable material alone or in combination with one or more other materials. In some embodiments, the cell bed20is defined by a monolithic paraffin body. In some embodiments, DNase and/or RNase inhibitors may be added to the fixative or other liquid solutions and/or paraffin that may improve future molecular testing.

In some embodiments, the base11is detachable, e.g., releasably attachable to a bottom portion10lof the tube10. This releasable attachment can be by any suitable attachment configuration including, for example, threaded attachment, bayonet or frictional fit, snap fit, hooks, VELCRO, adhesive attachment, frangible attachments, any of which may optionally also employ O-rings, compatible sealant, wax or grease or washers to promote a sufficient fluid-tight seal. For frangible attachments, the tube body10band/or base11can be integrally attached and configured to preferentially tear or detach about a defined zone when exposed to sufficient compressive, torsion or tensile forces.

The tube10can be provided in a sterile package40for onsite collection of a specimen from a patient. The term “onsite” refers to a collection location of a patient such as a surgical or biopsy room, doctor's office or hospital or laboratory site. The tube10can include a lid or cap12(FIG. 2C) that is provided in the package40or in a separate package. In other embodiments, non-sterile uses are contemplated.

The base11can be provided in the package40pre-assembled or pre-formed in the tube10and/or base11as shown. Alternatively, it may be provided separately for attachment at the use (collection) site (FIG. 11). As such, the cell bed20may be provided as a separate component in the package40or a separate package (held in a rigid container so as to protect the preformed shape). In other embodiments, the cell bed20is pre-assembled into the base11or bottom10lof the tube body10b. In some particular embodiments, the cell bed20can be formed in the tube10at the collection site if a suitably reliable press or formation system can be included in a collection kit.

In some embodiments, the cell bed20can be pre-formed in the tube10with a defined three-dimensional shape, in package40, so that the tube10is ready for use at a cell collection site. The package40can hold a single tube or multiple tubes10. The cap12(FIG. 2C) can be on the respective tube10to maintain the sterility until use.FIGS. 2A-2Cillustrate exemplary steps that can be carried out at a collection site to collect cells for subsequent evaluation. As shown inFIG. 2A, a sample with cells C is introduced into the internal cavity10cof the tube10and rest on the cell bed20. The cells C may comprise aspirated cells (unclotted) from a FNA using a needle75that is directly inserted into the tube10, in some embodiments.FIG. 2Billustrates that the cells C may then clot (onsite).FIG. 2Cillustrates that a supernatant, e.g., a solution of fixative liquid15that may comprise formalin or other suitable fixative material such as Zinc can be introduced into the tube10. Other fixatives may include, but are not limited to, saline, alcohols, acetone, mercury based reagents, and even media (Lennox broth, RPMI, etc.). The vessel10can be provided unfilled and a user can select the appropriate fixative or several or a particular type may be prepackaged in a kit which may be ordered for use. Any media used in the tube body10bshould be sterilized.

A lid12can be attached to the tube10and transported or stored. As shown, there may be undesired floating cells F in the solution above the clotted cells on the cell bed20.

The lid12can be a rigid closed lid that is attached after the fixative15is introduced. However, in some embodiments the lid12can include a liquid entry port to allow the liquid to enter while the lid remains on. The lid12can include a luer lock or luer slip connection fitting that engages with a male syringe luer lock or slip fitting to provide the liquid entry hat allows the liquid to be introduced through the port.

FIGS. 3A-3Fillustrate steps that can be carried out at a cytology laboratory or other suitable research or clinical laboratory.FIG. 3Ashows the tube10after centrifugation in a centrifuge200(FIG. 10). The centrifuge can be a standard laboratory centrifuge, typically a low speed centrifuge that permits the separation of the fixative from the cells and allows the cells to form a cell pellet P as is known to those of skill in the art. The centrifuge may be configured to process standard 50 mL or 100 mL conical tubes and the tube10can be placed therein alone or with an adapter. That is, the tube10may include a sleeve, adapter, or coupler or may have an external integrated size and/or design that allows it to be placed directly into the “bucket” or standard receptacle of the centrifuge.

The fixative liquid15is removed (e.g., the formalin is decanted) as shown inFIG. 3Band a rinse solution18can be added to the tube10as shown inFIG. 3C. The fixative liquid15can be removed and the rinse18added via any suitable technique that leaves the cells C and/or pellet P in the tube10including aspiration tubing, pipette withdrawal, decanting and the like. Typically, the supernatant should be aspirated gently with a vacuum rather than being decanted (which refers to tipped and poured) to minimize or reduce trauma to the cell pellet. As before, the rinse solution or other liquids can be removed or added with the lid12off as shown inFIG. 3Bor with the lid remaining on the tube using a liquid entry and/or retraction port. It is also contemplated that different lids having the same or different configurations may be used at different points in the process.

In some embodiments, a clot blot formed during the collection/post-collection can be used as a cap for a rinse vessel.

FIG. 3Dillustrates that the tube10with the rinse18can then be centrifuged.FIG. 3Eillustrates that the rinse18can be decanted or otherwise removed or withdrawn, leaving cells C on the cell bed20, typically in pellet form P.FIG. 3Fillustrates that a matrix material28can be added to form a cell block100. The term “matrix material” refers to a specimen-processing gel (that may be aqueous) that encapsulates and suspends histologic and cytologic specimens in a solidified medium. The matrix material28can include one or more of agar, agarose gel or “histogel” solid at ambient temperature, Methocell®, Matrix Gel®, OCT compounds, paraffin, denatured and non-denatured collagen, fibronectin, laminin, plasma and thrombin and other mixtures. Other matrices for cell immobilization can also be used. For a discussion of cell blocks and ethanol formalin fixative and other fixatives, see, e.g., Nathan et al.,Improved Preparation and Its Efficacy in Diagnosing Cytology, AmJ Clin Pathol, 2000; 1114, 599-606, the contents of which are hereby incorporated by reference as if recited in full herein.

FIG. 4Aillustrates that the base11and body of the tube10bare detached from each other, exposing the cell block100on the cell bed20. The cell block100with the cell bed20are removed from the base11as shown inFIG. 4B. The cell block100with cell bed20can be detached, removed or separated in any suitable manner including, for example, scraping, sliding or lifting (e.g., using a liner110,FIG. 4C).

FIG. 5illustrates the resulting cell block100(also termed cell disk) that includes the cell bed20ready for routine processing. The cell block100can be sliced or cut for preparing slides for staining or other diagnostic protocols. There may be an increased number of cells in the cell block or slices thereof that may promote diagnostic capability over smears alone.

FIG. 6Aillustrates the tube10with the base11detached from the tube body10b. In this example, the base11includes a raised center pedestal11pthat forms the inner surface11ithat holds the cell bed20. The perimeter of the pedestal11pcan include threads that engage an inner surface of the tube body or may frictionally engage the tube body using seals such as an O-ring(s) and the like (not shown). Typically, the base11with the cell bed20on the pedestal is attached to the tube body10band packaged for use at a collection site.

FIG. 6Billustrates that the tube10can include a spacer13that rises a distance above the lower portion of the base11.FIG. 6Balso illustrates that the base11can attach to the tube body10bat a location above the bottom of the base11. The spacer13can include a substantially planar or flat surface for holding the cell bed20. The base11and tube body10bcan threadably couple together. In the embodiment shown, the base11includes external threads34and the tube body10bincludes internal threads38. However, as noted above, other coupling configurations may be used.

FIG. 6Cillustrates that the base11can interchangeably attach to two different tube bodies10b1,10b2, and/or the different tube bodies can have different volumes. Thus, for example, the smaller tube body10b1can be used at the collection site and for transport to the cytology lab. The larger tube body10b2can be used at the cytology lab for processing in the centrifuge, for example. The same or differently configured caps or lids12may be used for each tube body10b1,10b2. In other embodiments, different volume tube bodies10b1,10b2can be provided in a package and selected for use at the collection site allowing for increased flexibility corresponding to the specimen type (e.g., urine, blood plasma or serum versus FNA).

FIG. 6Dillustrates that the tube body10bcan include several segments10b1,10b2that attach together to provide a different volumetric capacity. Thus, for example, one segment10b1can be attached and used with the cap or lid12at the collection site and for transport. The lid/cap12can be removed and the second body10b2can be attached to the first body10b1at the cytology lab and the stacked segments can define the tube body10bused for centrifugation. Where more than one tube body segment10b1,10b2is used, one or both can be detached from each other and/or the base11to expose the cell bed20with the cell block100for access/removal of the cell block100for subsequent processing and analysis.

FIG. 7Ais an exploded view of another example of a tube10. In this example, the base11has an annular open space33with female threads34that surrounds the internal surface11ithat holds the cell bed20. The female threads34matably engage external male threads38on the bottom portion of the tube10l.

FIG. 7B-7Eillustrate that the tube10can include top and bottom indicia12m,11mso that a user knows which end is “up” before using or opening. In some embodiments, a visually transparent window may be provided in the tube or cap or base or the device may be transparent or translucent. The cell bed20can have a substantially conical shape with the lowest peak facing the base11along an axially projecting centerline of the tube body10b. The outerwalls of the cell bed20wcan extend above the center portion of the cell bed20in a substantially straight vertical orientation so as to conform to the sidewalls of the tubular body10b. The sidewalls20wcan cover more than a major portion (e.g., greater than 50%) of the enclosed fluid cavity10c, leaving a minor portion of the sidewalls10wof the tube body10bbelow the cap12free of the cell bed material.

The tube10can have a defined capacity or volume. The tube10may have a volume or capacity between about 10 mL to about 200 mL, including about 20 mL, about 30 mL, about 40 mL, about 50 mL, about 60 mL, about 70 mL, about 80 mL, about 90 mL, and about 100 mL. The tubes10can be provided in different volumes/sizes for different applications. Where two segments10b1,10b2are used, one can have a volume between 10 mL to 25 mL and the other can have a volume between 25 mL and 100 mL, for example.

The lid12can also be threadably attached to an upper portion of the tube body10via threads12t,138. The base11and cap12can have a ledge39,139with a diameter that defines a tight fit with a receptacle of a standard centrifuge or with a standard tube, sleeve or other adapter allowing the tube10to be placed in a centrifuge for processing.

The cap and base ledges39,139can be configured to have the same outer diameter size. The outer perimeter of the ledge can include a pattern of circumferentially spaced apart recesses or grooves39g,139g. The ledges39,139can provide a resilient fit to provide for snug engagements using an overcoat, outerlayer or substrate of resilient material or just based on the groove configurations.

In some embodiments, the tube10is sized and configured as a 50 mL tube and can snugly engage a centrifuge receptacle without the use of an adapter or without a customized sleeve or other adapter.

FIGS. 8A and 8Billustrate that the tube10can be placed inside a larger (standard) tube50for standard centrifuge processing. The ledges39,139can snugly engage the sidewall of the tube50. The tube50can include a cap52and the lower portion54can include a center with a conical internal shape55. The tube10typically resides over the conical shape in the bottom portion of the standard tube50. However, the tube10can be placed above this location as well. The tube10can be slid in and out of the tube50for access.

FIGS. 9A and 9Billustrate another embodiment of a container10′ where the tube body10bcan releasably attach to an elongate body150which may be attached to a second cap12b. As shown, the elongate body150is in fluid communication with the tube fluid cavity10c. The second tube cap12bcan define a fluid port153that allows fluid from the elongate body150to be introduced or withdrawn. The tube body10bwith the second cap12bcan be configured to reside in a standard 50 mL tube50as shown. In other embodiments, the tube body10band second cap12bcan be configured to snugly reside in a centrifuge receptacle (e.g., bucket) without requiring the tube50. The elongate body150can be integral with the tube cap12b, e.g., a monolithic molded unitary component. In other embodiments, the elongate body can be configured to attach to the lid12bin other ways including threaded, snap fit and the like, while providing a fluid-tight seal.FIGS. 9C and 9Dillustrate a similar configuration of a container10′ with an alternate top configuration. In some embodiments, the elongate body150may be attached at the collection site as the original cap. In other embodiments, a cap or lid12such as that shown inFIG. 7Acan be used at the collection site and/or for shipment and transport and the second cap12bor elongate body150interchanged later for processing.

FIG. 10Ashows the tube10sitting over a spacer30snugly attached to and in a standard 50 mL tube50in preparation for centrifugation in a bucket210of the centrifuge200.FIG. 10Billustrates the tube10in the tube50in a standard swing bucket210of the centrifuge200.FIG. 10Cillustrates centrifugation with the tube10on its side (and horizontal) rather than angled so that the specimen is collected in a middle20c(FIG. 1B) of the cell bed20.

FIG. 11illustrates the components of a tube10according to some embodiments. As noted above, the cell bed20can be pre-formed and provided for assembly onsite or may be pre-attached to the base11and/or tube body10b. Typically, the base11with the cell bed20is attached to the tube body10band packaged for use at a collection site.

FIG. 12illustrates that the cap12′ can be configured to engage a leur-lock of a syringe120for introducing and/or removing different liquids.

FIGS. 13A and 13Cillustrate examples of a mold that has mold cavities20c1-20cn, that can concurrently form a plurality of cell beds20. Cell bed material, in liquid, solid, or semi-solid form, typically in flowable (fluid or gel) form can be introduced into the mold via one or more mold ports or through an open access region and molded into the pre-formed cell bed shape for use in the tube10.

In some embodiments as shown inFIG. 13B, a mechanical press251with shaped mold members253can be configured to enter the tube body10bwith an attached base11held in a lower holding member252to allow concurrent cell bed formation in multiple tubes10. The cell bed material20mcan be inserted in solid or liquid form, typically in a solid, but perhaps slightly heated form for aid in formation of the desired cell bed shape.

In other embodiments, as shown inFIG. 14, a respective cell bed20is formed directly in an attached base11and tube body10busing a shaped tamper tool260.

In some embodiments, the tubes10can be used to process cells for human or veterinary uses. In certain embodiments, the tubes10can be directed to preparation of cells for pathology review. While it is contemplated that the tubes10are particularly suitable for cells obtained by fine needle aspiration, it should be clear to one of skill in the art that cellular material captured by other means could also be collected and processed by the tubes10.

Cell material could also be collected by endoscopy, including but not limited to arthroscopy, bronchoscopy, colonoscopy, colposcopy, cystoscopy, ERCP (endoscopic retrograde cholangio-pancreatograthy), EGD (esophogealgastroduodensoscopy), endoscopic biopsy, gastroscopy, laparoscopy, laryngoscopy, proctoscopy and thoracoscopy. Cells could also be obtained from lavage procedures, including but not limited to bronchoalveolar, breast ductal, nasal, pleural, peritoneal, gastrointestinal, arthroscopic, and urinary bladder lavages. It is also contemplated that cells could be collected from catheters such as those used in infusion, cardiovascular, renal, bladder, urethral, hemodynamic monitoring, neurological, and other procedures which would be obvious to one of skill in the art. In some embodiments, cell samples can be from eye/cornea/globe aspirations, endocervical/ectocervical/endometrial curettages, cyst aspirations and urine. It is also contemplated that cell samples can be for xenografts from research and animal modeling as well as patient directed therapy.

The cells can be from washings and spontaneously exfoliated specimens including bronchial washings, bronchoalveolar lavage, sputum pleural fluid, pericardial fluid, peritoneal fluid, peritoneal washing, ovarian cyst fluid, synovial fluid, urine, brain cyst fluid, cerebrospinal fluid. The cells can be for RNA/DNA research or analysis and may include live cells. With the use of appropriate media, the tubes10can act as a small incubator to keep cells alive (at least for a short period of time). DNase/RNase inhibitors can be introduced to the media to also preserve DNA/RNA. As is known, fixation alone can help with DNA/RNA preservation.

In particular embodiments, the cell samples are from endocervical curettages (ECC). In the past, conventional practice when the first slide from the original paraffin block is essentially noncontributory, is to take the fluid remaining in the specimen jar and perform a “ThinPrep” on it. These typically have many cells (squamous and glandular) but there is no architecture. Rather than place the minimal slime usually present in a specimen jar in a cassette, it is contemplated that those cells can be put in the tubular body10b(or150,FIG. 9A, 15A) at the collection site. This should give better initial yield with architecture present and a source for immunos. This latter may not be inconsequential. For example, when the ECC contains a minute fragment of small cells with high N/C ratios, it is hard to discern whether they are not relevant (being potentially from the lower uterine segment) or clump of HGSIL cells. A single immuno-p16—can be very useful in this scenario. The term “immuno” and plurals thereof refer to immunoperoxidase studies and include antibodies targeting specific epitopes to aid in tumor/disease differentiation. Also known as (although technically incorrect) immunohistochemistry: p16 is a protein/antigen with the p16 antibody in a cell having clinical significance.

FIGS. 15A-15Iillustrate another embodiment of a container10″ similar to that shown inFIGS. 9A and 9B. In this embodiment, no external tube is required for centrifuge processing. As shown, the container10″ includes a tube body10bholding the cell bed20and a base11. The container10″ also includes an elongate body150that can attach to the tube body10b. The elongate body150can include a first end with a lid155and a second opposing end157that is open. As shown inFIG. 15A, the elongate body150can include a desired solution, such as saline or formalin15.FIG. 15Billustrates that, as before, a FNA sample can be inserted onto the cell bed20. One or more needles from other passes obtained from the target tissue can be rinsed in the chamber of the elongate body150. This is in contrast to a jar filled with saline used conventionally. As shown inFIGS. 15C and 15D, the tube body10band base11can be attached together after the pellet P is dried allowing the tube body10bwith cell bed20and pellet P to be inverted to attach to the open end157of the elongate body150. The open end of the elongate body157can be sized and configured to be substantially the same as the size of the upper end of the tube body. The elongate body157can taper to a large size away from the open end as shown. The tube body10bcan engage the open end of the elongate body157in a fluid tight manner using appropriate seals, threads, frictional fits and the like.

FIG. 15Eshows that the container10″ can be inverted so that the cell bed20is in the lower portion of the container10″. The container10″ can be centrifuged at an appropriate revolution per minute and duration (e.g., about 2000 rpm for about 5 minutes) to form a combined pellet Pc (FIG. 15F) on the cell bed20(combined from cells collected from the rinse solution and the FNA direct deposit).FIG. 15Gillustrates that the supernatant can be aspirated as is conventional using a vacuum leaving only a minimal amount of fixative15(e.g., formalin or saline) in the tube body10babove the cell bed20so as to not disrupt the cell pellet P. The aspiration can be carried out by first removing lid155or by using a sealed port in the lid (not shown). As shown inFIG. 15H, the elongate body150can then be removed. A liquid matrix28can be added to the tube body10band cells resuspended.FIG. 15Iillustrates that the cell disk or block100with the cell bed20can be removed from the tube body10busing, for example, a plunger300. The cell block100can be processed as a histology specimen or other desired specimen.

FIG. 16is a flow chart of exemplary operations that can be used to carry out embodiments of the invention. A cell sample can be placed in a tube having a cell bed (block350). The sample is centrifuged while in the tube (block360). A cell block is removed from the tube with the cell bed and solidified matrix holding the cells (block370).

The cell sample can be placed with a needle with a fine needle aspirate sample directly in the tube (block352). Liquid matrix material (e.g., specimen-processing gel that encapsulates and suspends histologic and cytologic specimens in a solidified medium) can be added to the tube after the centrifuging and the matrix material with the cells distributed therein can be solidified to form the cell block (block365). The cell sample can be collected in the tube at a patient collection site and the tube can be transported to a cytology lab for processing (block355).

The base of the tube can be separated from the tube body to expose the cell block and allow the cell block to be removed with the cell bed (block368).

In some embodiments, generally summarized, rinses can be performed in the collection vessel10and the dedicated clot blot(s) can be generated as discussed above. The clot blot can be used as a cap for the rinse vessel. The entire apparatus can be inverted (clot block side down) and centrifuged. The supernatant can be aspirated leaving a clot blot with overlaying precipitated materials (button). A desired amount (not typically calculated precisely, but roughly, about 1:1/v:v to the button) of HistoGel can be pipetted onto the button and immediately spun again in a centrifuge. This time the collection vessel is not needed. This allows the HistoGel to permeate the cells and polymerize. Again, this step is carried out quickly to prevent polymerization before the centrifugation. This leaves the cells within a matrix of HistoGel. This action can prevent loss of cells in downstream processing procedures.

EXAMPLES

FIGS. 17-27are digital images of an exemplary tube10and exemplary processing that can be carried out using the tube (the centrifuge operations were shown above with respect toFIGS. 10A-10C).FIG. 17illustrates a FNA expelled directly from a needle into the cell bed20of the base11(which is attached to the tube body10b). As shown inFIG. 18, the cell sample C can settle in the middle of the cell bed20and/or base12due to the substantially conical or furstoconical shape of the cell bed20. The sample can be allowed to dry and/or coagulate.

The supernatant (formalin) is aspirated and discarded being careful not to disrupt the pelleted sample P as shown inFIG. 19.FIG. 20illustrates the tube10with a sample pellet P. Some supernatant may remain but does not interfere with subsequent processing.

After placing the tube10in the tube50, and centrifuging (as described above), liquid histogel (melted agarose) can be pipetted up and down in the tube10as shown inFIG. 21using pipette299to resuspend the sample in the histogel. About 30 μL of agarose may be used for this resuspension although other amounts may also be appropriate.FIG. 22illustrates that the suspension can be carried out relatively rapidly so that the agarose does not solidify.FIG. 23shows the tube with the suspended sample in a low temperature freezer (e.g., about −20 C.) to facilitate solidification, which can occur in between about 2-5 minutes.

FIG. 24illustrates the cell block100(e.g., the cell bed20and the solid histogel with cells) being removed from the tube10band/or base11.FIG. 25shows that the cell bed20may attach to the inner wall of the tube body10band may need a plunger or other push member300to push the cell bed with the cells, e.g., cell block100from the tube body (typically out the top, pushing against the cell bed20rather than the cells/histogel (agarose) mixture100.

FIG. 26illustrates the cell block100with cell bed20placed on routine tissue paper. As is known to those of skill in the art, it is standard practice to wrap the sample100in tissue paper in preparation for tissue processing for small histology specimens.FIG. 27illustrates the tissue paper wrapped cell block100with cell bed20in a standard histology cassette310. The darker dots inside the wrapped cell block100are cells C.

Although shown as a manual operation, it is contemplated that machines may be used to automatically carry out certain of the above steps.

Table 1 below is a list of 15 cases of data comparing an exemplary collection vessel with the standard methods. All cases were graded from1(poor) to2(adequate) to3(superior). The first row is the diagnosis. The 2nd row is the paired smear quality (DQ and H&E). The CBCS is the remnant material left on the traditional clotting slide. The clot blot is the standard method compared to the cell disk method (CD). Last level was the last recut level on the FFPE block. The method was overall graded for superiority. As seen, the standard method was never superior to the new CD method but equivocal in some cases.