WO2019083772A1

WO2019083772A1 - Containers suitable for cell pathology processing and associated cell processing methods

Info

Publication number: WO2019083772A1
Application number: PCT/US2018/056015
Authority: WO
Inventors: John David DRUGA, Sr.; Philip Jayson BROWN; Aaron VAN GORKOM
Original assignee: Wake Forest University Health Sciences
Priority date: 2017-10-25
Filing date: 2018-10-16
Publication date: 2019-05-02

Abstract

Containers with a tubular member having opposing first and second end portions and an open fluid through channel. The second end portion has a radially extending lip. The containers include a base releasably and sealably attachable to the tubular member. The base includes a medial portion with a shaped recess having a closed bottom surface and at least one drainage aperture above the closed bottom surface of the shaped recess. When assembled to the tubular member, the radially extending lip sealably covers the at least one drainage aperture.

Description

CONTAINERS SUITABLE FOR CELL PATHOLOGY PROCESSING AND

ASSOCIATED CELL PROCESSING METHODS

Related Applications

[0001] This application claims the benefit of and priority to U.S. Provisional

Application Serial No. 62/576,891, filed October 25, 2017, the contents of which are hereby incorporated by reference as if recited in full herein.

Field of the Invention

[0002] This invention relates to containers particularly suitable for cell pathology, cell collection and/or cell processing.

Background of the Invention

[0003] 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.

[0004] 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 or other samples. This cell sample 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

[0005] Embodiments of the invention provide containers that can directly receive cells from a FNA or other sources at a collection site and can provide direct access to a solid cell pellet by simply removing a base with an interior shaped recess that provides a cell bed.

[0006] 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 solid cell pellet with suspended cells, optionally in the form of a rounded (semi-spherical) solid cell pellet.

[0007] Embodiments of the invention are directed to containers that include a tubular member with opposing first and second end portions and an open fluid through channel. The the second end portion has a radially extending lip. The containers also include a base that is releasably and sealably attachable to the tubular member. The base includes a medial portion with a shaped recess having a closed bottom surface and at least one drainage aperture above the closed bottom surface of the shaped recess. When assembled to the tubular member, the radially extending lip sealably covers the at least one drainage aperture.

[0008] The base can also include an outer wall and an inner wall. The outer wall can include threads that attach to mating threads of the tubular member to releasably attach to the tubular member. The inner wall can have a first wall segment forming an upwardly extending wall of the shaped recess and a second wall segment facing the outer wall. The first inner wall segment can be joined to the second inner wall segment by a peak wall segment. The at least one drainage aperture can reside at least partially on the peak wall segment.

[0009] The lip can extend at least partially about a circumference of the fluid through channel.

[0010] The lip can extend about an entire circumference of the fluid through channel and can have a free inner end that tapers down and inward from an outer wall of the tubular member.

[0011] The tubular member can have a monolithic molded polymer body with the lip moldably integral therewith. [0012] The second end portion of the tubular member can have threads that extend at least partially below an inner end of the lip.

[0013] The at least one drainage aperture can be a plurality of circumferentially spaced apart drainage apertures.

[0014] The at least one drainage aperture can have an elongate radially extending shape and an end facing the outer wall can have a greater diameter than an end facing the shaped recess.

[0015] The base can also include: an outer wall; an inner wall; and an annular cavity between the outer wall and the inner wall. The outer wall can surround the inner wall and the inner wall can include first and second radially spaced apart wall segments joined by a peak wall segment, the first wall segment defining an interior wall of the shaped recess, and wherein the at least one drainage aperture can extend radially at least partially across the peak segment.

[0016] The second end portion of the tubular member can extends a distance into the annular cavity.

[0017] The shaped recess can have an aspect ratio h/w of between 1 and 1.5.

[0018] The tubular member and attached base can provide a liquid volumetric capacity that is between about 10 mL to about 100 mL.

[0019] The at least one drainage aperture can include a plurality of circumferentially spaced apart drainage apertures and the drainage apertures can have an elongate radially extending shape.

[0020] The base can have an axially extending centerline and the bottom of the shaped recess can have a radius of curvature "R" that is between 3-6 mm.

[0021] The base can have a plurality of external "V" shaped partition walls providing a planar bottom surface for the base.

[0022] The container can include a cap configured to attach to the first end portion of the tubular member. The tubular member can have a length that is greater than that of the base.

[0023] Other aspects are directed to methods of collecting and processing a biosample.

The methods include: (a) providing a container with a tubular member sealably attached to a releasable base, the base comprising at least one drainage aperture above a shaped recess in the base, the shaped recess forming a cell bed, the tubular member comprising a lip that sealably covers the at least one drainage aperture, and the container comprises a biosample comprising cells that reside in the shaped recess; (b) centrifuging the biosample in the tubular container so that cells from the biosample deposit as a pellet against the cell bed; (c) inserting a liquid material in the tubular container above the cell bed; (d) forming a solid cell pellet holding distributed cells therein; (e) loosening the tubular member from the base to unseal the lip from the at least one drainage aperture; (f) draining liquid from the base via at least one drainage aperture above the cell bed in response to the loosening action; (g) detaching the base from the tubular member; and then (h) removing the solid cell pellet from the base.

[0024] The draining can include flowing liquid in linear parallel streams from circumferentially spaced apart drainage apertures in the base.

[0025] The biosample can include cells from a fine needle aspirate tissue sample.

[0026] Other aspects are directed to tubular members for a specimen processing container. The tubular members include: opposing first and second end portions and an open fluid through channel. The second end portion includesexternal threads on a wall thereof and a moldably integral, radially extending internal lip that extends at least partially about a circumference of the fluid through channel.

[0027] The lip can extends about an entire circumference of the fluid through channel and can have a free inner end that tapers down and inward from the wall of the tubular member.

[0028] Still other embodiments are directed to a base for a cell processing container.

The base includes an outer wall and an inner wall spaced apart by an annular cavity. The outer wall can include internally extending threads. The inner wall can have a first wall segment forming an upwardly extending wall of the shaped recess and a second wall segment facing the outer wall. The first inner wall segment can be joined to the second inner wall segment by a peak wall segment. The base can also include a plurality of circumferentially spaced apart drainage apertures residing at least partially across the peak wall segment.

[0029] The at least one drainage aperture can have an elongate radially extending shape, and wherein an end facing the outer wall has a greater diameter than an end facing the shaped recess.

[0030] The shaped recess can have an aspect ratio h/w of between 1 and 1.5.

[0031] The base can have an axially extending centerline and the bottom of the shaped recess has a radius of curvature "R" measured from the centerline that is between 3-6 mm.

[0032] The base can include a plurality of external "V" shaped partition walls, each

V-shaped wall extending about one of the drainage apertures and the V-shaped partition walls have a planar external bottom surface providing a planar bottom surface for the base. [0033] The shaped recess can have a concave middle portion and/or a middle portion with a substantially conical shape that defines a closed solid cell bed surface.

[0034] The biosample can include cells from a fine needle aspirate that can be injected directly from a needle holding the aspirate into the container.

[0035] Other embodiments are directed to containers. The containers can have a tubular body having an open through channel and open opposing first and second end portions. The second end portion can have a radially extending lip that circumferentially extends about the open through channel.

[0036] A base with one or more drain apertures attaches to the tubular body and the at least one drainage aperture resides above a shaped recess with a closed bottom is removably attached to the second end portion of the tubular body with the lip sealing the drain apertures. The base can provide an internal solid cell bed surface.

[0037] The base can include an annular recess between an outer wall and an inner wall. The outer wall surrounds an inner wall with first and second radially spaced apart wall segments joined by a peak wall segment, the first wall segment facing the outer wall and the second wall segment defining an interior wall of the shaped recess over a closed bottom of the shaped recess. The at least one drainage aperture can extend radially at least partially across the peak segment.

[0038] The container can have a volume that is between about 10 mL to about 100 mL.

[0039] The tubular member can have threads on upper and lower portions thereof, the lower portion can be configured to threadably attach to the outer wall of the base.

[0040] The tubular body can be sterile and configured to hold human or animal cell samples.

[0041] Embodiments of the invention provide containers 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.

[0042] 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.

[0043] It is noted that aspects of the invention described with respect to one embodiment, may be incorporated in a different embodiment although not specifically described relative thereto. That is, all embodiments and/or features of any embodiment can be combined in any way and/or combination. Applicant reserves the right to change any originally filed claim or file any new claim accordingly, including the right to be able to amend any originally filed claim to depend from and/or incorporate any feature of any other claim although not originally claimed in that manner. These and other objects and/or aspects of the present invention are explained in detail in the specification set forth below.

[0044] Other systems and/or methods according to embodiments of the invention will be or become apparent to one with skill in the art upon review of the following drawings and detailed description. It is intended that all such additional systems, methods, and/or devices be included within this description, be within the scope of the present invention, and be protected by the accompanying claims.

Brief Description of the Drawings

[0045] The patent or application file contains at least one drawing executed in color.

Copies of this patent or patent application publication with color drawings will be provided by the Office upon request and payment of the necessary fee.

[0046] Other features of the present invention will be more readily understood from the following detailed description of exemplary embodiments thereof when read in conjunction with the accompanying drawings.

[0047] FIG. 1 is an exploded top perspective view of an example container according to embodiments of the present invention.

[0048] FIG. 2 is an assembled section view of the container shown in FIG. 1 according to embodiments of the present invention.

[0049] FIG. 3 is a side assembled view of the example container of FIG. 1.

[0050] FIG. 4 is a section view taken along line 4-4 in FIG. 3.

[0051] FIG. 5A is an exploded bottom perspective view of the example container shown in FIG. 1.

[0052] FIG. 5B is a partial side view of the container showing fluid flowing from the base in response to the base being loosened to allow fluid drainage according to embodiments of the present invention.

[0053] FIG. 5C is a partial side view of the container, with the cap off and the base attached, inverted to pour fluid out during processing according to embodiments of the present invention. [0054] FIG. 5D is an enlarged side perspective view of the container with liquid flowing from the drainage apertures while the base is still attached to the tubular member according to embodiments of the present invention.

[0055] FIG. 6 is a side view of a tubular member of the example container shown in

FIG. 1 according to embodiments of the present invention.

[0056] FIG. 7 is a section view of the tubular member taken along line 7-7 in FIG. 6.

[0057] FIG. 8A is an enlarged partial view of the lower portion of the tubular member shown in FIG. 7.

[0058] FIG. 8B is a greatly enlarged partial view of the base and lip of the container shown in FIG. 1 according to embodiments of the present invention.

[0059] FIG. 9 is a side view of the base of the example container shown in FIG. 1 according to embodiments of the present invention.

[0060] FIG. 10 is a section view of the base taken along line 10-10 in FIG. 9.

[0061] FIG. 11 is a section view of the base taken along line 11-11 in FIG. 9.

[0062] FIG. 12 is a top, side perspective view of the base shown in FIG. 9.

[0063] FIG. 13 is a top view of the base shown in FIG. 9.

[0064] FIG. 14 is an enlarged view of a drain aperture shown in FIG. 13.

[0065] FIG. 15 is a bottom view of the base shown in FIG. 9.

[0066] FIG. 16 is a section view of the base taken along line 16-16 in FIG. 15.

[0067] FIG. 17 is a flow chart of exemplary operations that can be used to process cells according to embodiments of the present invention.

[0068] FIGs. 18A-18D are digital photographs of an example cell pellet formed using the device of FIG. 1.

[0069] FIG. 19A is a cell pellet prepared using the device of FIG. 1 according to embodiments of the present invention.

[0070] FIG. 19B is a microscope slide view of the cell sample from the cell pellet shown in FIG. 19A according to embodiments of the present invention.

[0071] FIG. 19C is a cell pellet prepared using a conventional method and container.

[0072] FIG. 19D is a microscope slide view of the cell sample from the cell pellet of

FIG. 19C.

[0073] FIG. 20 is a side perspective view of a base of the container of FIG. 1 that can be used to prepare a cell pellet according to standard processing methods according to embodiments of the present invention. [0074] FIG. 21A and 21B are top perspective views of a centrifuge that can hold container such as that shown in FIG. 1 for processing according to embodiments of the present invention.

Detailed Description of Embodiments of the Invention

[0075] The present invention now is described more fully hereinafter with reference to the accompanying drawings, in which embodiments of the invention are shown. This invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein; rather, these embodiments are provided so that this disclosure will be thorough and complete, and will fully convey the scope of the invention to those skilled in the art.

[0076] 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.

[0077] The terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. As used herein, the singular forms "a", "an" and "the" are intended to include the plural forms as well, unless the context clearly indicates otherwise. It will be further understood that the terms "comprises" and/or "comprising," when used in this specification, specify the presence of stated features, integers, steps, operations, elements, and/or components, but do not preclude the presence or addition of one or more other features, integers, steps, operations, elements, components, and/or groups thereof. As used herein, the term "and/or" includes any and all combinations of one or more of the associated listed items. As used herein, phrases such as "between X and Y" and "between about X and Y" should be interpreted to include X and Y. As used herein, phrases such as "between about X and Y" mean "between about X and about Y." As used herein, phrases such as "from about X to Y" mean "from about X to about Y."

[0078] Unless otherwise defined, all terms (including technical and scientific terms) used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. It will be further understood that terms, such as those defined in commonly used dictionaries, should be interpreted as having a meaning that is consistent with their meaning in the context of the specification and relevant art and should not be interpreted in an idealized or overly formal sense unless expressly so defined herein. Well-known functions or constructions may not be described in detail for brevity and/or clarity.

[0079] It will be understood that when an element is referred to as being "on",

"attached" to, "connected" to, "coupled" with, "contacting", etc., another element, it can be directly on, attached to, connected to, coupled with or contacting the other element or intervening elements may also be present. In contrast, when an element is referred to as being, for example, "directly on", "directly attached" to, "directly connected" to, "directly coupled" with or "directly contacting" another element, there are no intervening elements present. It will also be appreciated by those of skill in the art that references to a structure or feature that is disposed "adjacent" another feature may have portions that overlap or underlie the adjacent feature.

[0080] Spatially relative terms, such as "under", "below", "lower", "over", "upper" and the like, may be used herein for ease of description to describe one element or feature's relationship to another element(s) or feature(s) as illustrated in the figures. It will be understood that the spatially relative terms are intended to encompass different orientations of the device in use or operation in addition to the orientation depicted in the figures. For example, if the device in the figures is inverted, elements described as "under" or "beneath" other elements or features would then be oriented "over" the other elements or features.

Thus, the exemplary term "under" can encompass both an orientation of over and under. The device may be otherwise oriented (rotated 90 degrees or at other orientations) and the spatially relative descriptors used herein interpreted accordingly. Similarly, the terms "upwardly", "downwardly", "vertical", "horizontal" and the like are used herein for the purpose of explanation only unless specifically indicated otherwise.

[0081] It will be understood that, although the terms first, second, etc. may be used herein to describe various elements, components, regions, layers and/or sections, these elements, components, regions, layers and/or sections should not be limited by these terms. These terms are only used to distinguish one element, component, region, layer or section from another region, layer or section. Thus, a first element, component, region, layer or section discussed below could be termed a second element, component, region, layer or section without departing from the teachings of the present invention. The sequence of operations (or steps) is not limited to the order presented in the claims or figures unless specifically indicated otherwise.

[0082] The term "about" means that the recited number or value can vary by +/- 20%. [0083] The term "biosample" refers to tissue, blood or other solid or liquid samples that have cellular material and may be particularly suitable for human or animal biosamples. 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.

[0084] 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.

[0085] 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.

[0086] Embodiments of the invention may be suitable for veterinarian use, medical human use or research.

[0087] 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).

[0088] Turning now to the figures, FIGS. 1-4 illustrate a specimen container 10. The container 10 can include a tubular member 20 that can sealably and releasably couple to a base 30 and can also include a cap 40. The base 30 can include at least one drainage aperture 32, shown as a plurality of circumferentially spaced apart apertures 32.

[0089] The base can include an inner wall 35 and an outer wall 37. The inner wall 35 can surround a shaped recess 33 with a closed floor 33f. The shaped recess 33 can form a cell bed. The outer wall 37 can have a greater height Ή" than a maximal height of the inner wall 35. The inner surface of the outer wall 37 can be threaded. The inner wall 35 and the outer wall 37 can be spaced apart by an annular cavity 39 that can surround the internal shaped recess 33 with a closed floor 33f. The base 30 can comprise external cavities 139 and shaped wall segments 140 (FIGS. 5, 9, 15, for example). The at least one drainage aperture 32 can reside on the inner wall 35 above the closed bottom 33b as will be discussed further below. The base 30 can have an external bottom 30b that is configured to sit flat on a support surface. The base wall segments 140 can have planar bottom surfaces 140b to provide the base bottom configuration and provide a partition wall or "fin" about one or more of the at least one drainage aperture 32.

[0090] The tubular member 20 can have an outer wall 22 that can comprise threads 23 that can engage the threads 39 on the base 30. As shown, the tubular member 20 comprises male threads as the threads 23 and the base comprises mating female threads as the threads 39 but the reverse can be true. For example, the outer surface of the outer wall can comprise male or female threads and the tubular member 20 can have an inner surface that engages those threads. Also, it is contemplated that frictional or other sealable attachments can be used to sealably attach the base 30 to the tubular member 20. The sealable and 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 tubular member 20 and/or base 30 can be integrally attached and configured to

preferentially tear or detach about a defined zone when exposed to sufficient compressive, torsion or tensile forces.

[0091] The tubular member 20 can have a lip 25 that can cover and sealably engage the at least one drain aperture 32 in the base 30.

[0092] The tubular member 20 can be a molded monolithic polymeric member and the lip 25 can be integrally formed thereon.

[0093] The tubular member 20 can have a height (i.e., length) that is greater than the height (i.e., length) of the base 30, typically by a 30-100% greater length. In some embodiments, the base 30 can have a height "H" associated with the outer wall 37 that is about 30 mm, while the assembled tube 20 with the base 30 can have a height that is about 1 10 mm.

[0094] The tubular member 20 has an open through channel 26. The lip 25 can cover, typically abut, a perimeter surface 32p (FIG. 14) adjacent to and surrounding the at least one drainage aperture 32 to seal that respective at least one drainage aperture 32 when properly assembled. The lip 25 can have sufficient rigidity to have a self-supporting shape with the lip 25 extending inward from the wall 22 of the tubular member 20 when apart from the base 30 (FIG. 8A) . The lip 25 can have sufficient flexibility to flex in response to contact with the seal surface 25s of the base 30 (FIG. 8B) adjacent the at least one drainage aperture 32. The lip 25 can optionally be arcuate, semi-circular or circular and extend circumferentially about a portion or an entire inner perimeter 26p of the channel 26 at the lower portion of the through channel 261. The lip 25 can be discontinuous about the perimeter of the tubular member 20. The lip 25 can have a constant inward extent or different inward extents about the circumference of the tubular member 20.

[0095] The lip 25 can angle downward toward its inner end away from the outer wall

22 as shown in FIG. 8A and FIG. 8B, for example. The lip 25 can have a radially extending extent Di, measured from the inner surface of the wall 22 of the tubular member 20 to the tip of the lip 25t, that is between 1 mm and 10 mm, typically between about 1.5 mm and about 5 mm, and more typically about 4 mm to about 5 mm, in some particular embodiments.

[0096] The upper portion of the open channel 26u can have a greater diameter than the bottom portion of the open channel 26/.

[0097] Referring to FIGS. 4, 5A, and 6-8A and 8B, the tubular member 20 can have a first end portion 28 that extends below the lip 25. As shown in FIG. 8A, the lip 25 can taper inward and downward from the outer wall 22 at an angle "β" relative to the outer wall 22 at the first end 28, where β can be between 30-60 degrees.

[0098] As shown in FIGS. 2, 4 and 8B, the first end 28 of the tubular member 20 can extend into the annular cavity 39 and can terminate a distance above the closed end 39c of the annular cavity 39.

[0099] Referring to FIG. 8B, when in a sealing configuration with the base 30, the lower surface of the lip 25b from the hinge joint 25h to a medial or forward segment adjacent the tip 25t can reside above the peak segment 35p of the inner wall 35. The drainage aperture 32 and/or peak segment 35p can reside a distance D₂ away from the inner surface 28i of the first end 28 of the tubular member between 1 mm and 5 mm, more typically in a range of about 2 mm and about 3 mm.

[00100] FIG. 5B and 5D show fluid flowing from the base 30 in response to the base 30 and tubular member 20 being loosened to remove or loosen the seal provided by the lip 25 (i.e., the "lip seal" 25s (FIG. 2)) and allow fluid drainage via the at least one drainage aperture 32 according to embodiments of the present invention. The base 30 and/or tubular member 20 can be rotated a partial turn or one or more turns to loosen the seal provided by the lip 25 over the at least one drainage aperture 32 which can then rotated in the opposite direction to be tightened again for resealing.

[00101] FIG. 5C is a partial side view of the container 10 sans cap 40 with the base 30 attached and the container partially inverted to pour fluid out of the container 10 during processing according to embodiments of the present invention. [00102] FIGs. 5C and 5D show that the shaped wall segments 140 (e.g., wall partitions or fins) can cooperate with the drainage apertures 32 to form parallel linear fluid flow streams 139f of liquid L that may allow controlled draining while the cell sample collects in the medial concave region of the base 30 and with the base 30 still attached to the tubular member 20 according to embodiments of the present invention. The controlled flow can avoid or inhibit removing cells from the cell bed during the draining.

[00103] Referring to FIGS. 10, 11, 12, 13 and 16, the inner wall 35 of the base 30 can have first and second wall segments 35i, 35o. The first wall segment can be an inner wall segment 35i that forms a solid upwardly extending wall around the shaped recess 33. The second or outer wall segment 35o can face the annular cavity 39 and the outer wall 37. The second wall segment 35o can be substantially parallel to the outer wall 37 (FIG, 4, for example). The word "substantially" with respect to this feature means within about 10% of parallel orientations with respect to each other. The inner wall segment 35i can be curvilinear and taper down to the closed bottom 33b of the shaped recess 33.

[00104] The first or outer wall segment 35o and the inner wall segment 35i can be joined at a peak or shoulder segment 35p with a maximal height that can be less than the height of the outer wall 37. The inner wall segment 35i of the inner wall 35 can have a first segment 35t that tapers down from the peak 35p at an angle measured from the axially extending centerline C/L, that is between 5 degrees and 10 degrees, such as about 8 degrees, then can merge into a lower wall segment 35/ with a radius of curvature R that extends to the closed bottom 33b of the shaped recess 33.

[00105] Still referring to FIG. 16, the shaped recess 33 can have a radius of curvature measured relative to the axially extending centerline C/L that is in a range of about 1 mm to about 10 mm, more typically between about 3-6 mm, which can form a round, substantially spherical or semi-spherical shaped solid cell pellet when processed in the container 10 as shown by the broken circular perimeter in FIG. 16.

[00106] FIGs. 18A-18C are digital photographs of an example shaped cell pellet P formed in the base 30. As shown, the formed cell pellet P can comprise a body with an arcuate body shape, with an arc extent that can be between 180-360 degrees. The cell pellet P can have an arcuate outer perimeter with a radius of curvature between about 1 mm to about 6 mm, more typically between about 3-6 mm, such as about 3 mm, about 3.5 mm, about 4 mm, about 4.5 mm, about 5 mm, about 5.5 mm and about 6 mm, which can be measured when removed from the base 30 and held on a flat spatula. The cell pellet P can have a substantially spherical or semi-spherical shaped (typically a soft or gel-like solid) body that has increased cell concentration per volume and/or a smaller surface area than a cell pellet formed using known conventional devices.

[00107] Referring to FIG. 11, the shaped recess 33 can have a height "h", measured below the peak 35p of the inner wall at the inner wall segment 35i that is in a range of about 10 to about 15 mm and a width "w" that is in a range of about 9 mm to about 12 mm. In some embodiments, the shaped recess 33 can have an aspect ratio, h/w that is between 1 and 1.5.

[00108] Referring to FIGS. 12, 13 and 14, the at least one drainage aperture 32 can comprise ten equally spaced apart drainage apertures 32. However, more than ten or less than ten drainage apertures 32 may be used. Also, the drainage apertures 32 can be asymmetrically positioned and one or more may have different shapes and/or sizes than others. The drainage apertures 32 can be configured as elongate slots that extend radially out away from the shaped recess 33. As shown, the drainage apertures 32 can reside at least partially on or adjacent the peak segment 35p of the inner wall 35. The at least one drainage aperture 32 can extend laterally or radially across the peak segment 35p and down toward the outer wall segment 35o of the inner wall 35. As shown in FIG. 14, the at least one drainage aperture 32 can have an inner end portion 32i on the peak segment 35p can have a smaller diameter than an outer end portion 32o that can face the outer wall 37.

[00109] In some embodiments, the base 30 can comprise drainage apertures in the outer wall 37 and/or bottom wall above the annular cavity 39 (FIG. 10).

[00110] FIGs. 12 and 15 illustrate that, when viewed from the bottom, the wall segments 140 can have a "V" shaped "fin" configuration surrounding a medial open segment of the base 30m under the closed bottom 33b of the shaped recess 33. As shown in FIG. 15, each "V" shaped wall segment 140v can extend about one drainage aperture 32 and partition a corresponding cavity 139. There can be five V-shaped wall segments 140 that are circumferentially spaced apart, each extending about a corresponding drainage aperture 32. One drainage aperture 32 can extend between adjacent pairs of the V-shaped wall segments.

[00111] The wall segments 140 can be circumferentially spaced apart about the medial segment of the base 30m and can have greater rigidity than the medial segment of the base 30m. The medial external segment of the base 30m corresponds to the floor 33f of the shaped recess 33 (FIG. 15).

[00112] Surprisingly, the base 30 with the shaped recess 33 can provide a round cell pellet with an increased concentration of cells S that are primarily clustered within a diameter of a single view V of a microscope for easy viewing in a microscope view of a slide holding a cell sample S taken from the cell pellet P as shown in FIG. 19B. The view V in FIG. 19B is of a sample cell pellet P in the pouch 200 in FIG. 19A and has a compact and relatively dense concentration of cells with a perimeter with an arcuate shape over a major extent thereof and an associated radius of curvature(s) R. In contrast, the cell pellet Pc produced under a current known method and container results in a more distributed cell pellet Pc in the pouch 200 that takes up a larger area of the holding pouch or packet 200 and has a more distributed shape which requires many changes in slide position to view the sample under the microscope view V as shown in FIG. 19D. The sample S from the cell pellet Pc has a cell concentration that is distributed and 1.5X-2X larger on a respective slide than the cell pellet P. Stated differently, 80-95% of the cells of the cell pellet P can be within a diameter of a single slide field of view (which can optionally be between 2-25 mm or between 3-10 mm) as shown in FIG. 19B.

[00113] After processing, such as adding agents to clot/solidify the cells into the cell block, the tube 20 can be loosened or detached from the base 30 whereby the lip 25 no longer seals the at least one drainage aperture 32, allowing excess liquid to drain from the base 30.

[00114] As shown in FIG. 18A, when the base 30 is detached, the detached base 30 can expose or provide direct access to a suspension of a compact solid (typically round) cell pellet P. The top of the cell pellet P can reside under the peak segment 35p of the inner wall 35 (FIG. 18D). The cell pellet P can be easily be dislodged using a suitable tool such as a spatula or merely by turning the base upside down into a cassette thereby separating the solid cell pellet from the base 30 without physical trauma to the cells therein.

[00115] The shaped recess 33 of the base 30 can provide an inert cell bed. The cells can directly contact the polymeric material of the base or the shaped recess can have a surface with any suitable coating. The term "inert cell bed" refers to a solid material that can support processed cell material, typically in a block form, while preserving the cells, typically without chemically interacting with the cells. Post collection and after processing, the cell pellet P with patient cells can be removed, substantially intact, e.g., scraped, pushed, poured or otherwise disengaged from the base 30 substantially without disrupting the collected cells thereon for cell evaluation.

[00116] In some embodiments, the tubular member 20, base 30 and cap 40 can comprise a molded polymeric material that is sterilized for use. The tubular member 20, base 30 and/or cap 40 may also comprise other suitable materials, including, for example, ceramic, silica or glass. [00117] In some embodiments, DNase and/or R ase inhibitors may be added to a fixative or other liquid solutions while the cells are in the base 30 when attached to the tubular member 20 or when detached that may improve future molecular testing.

[00118] The container 10 can be provided in a sterile package 140 (shown by the broken line box in FIG. 3) for 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. In other embodiments, non-sterile uses are contemplated. The package 140 can hold a single specimen container or multiple such containers 10. The cap 40 can be on the respective tube 10 to maintain the sterility until use.

[00119] In some embodiments, a sample with cells can be introduced into the container 10 to rest on the internal surface of the solid floor 33f of the shaped recess 33. The cells may comprise aspirated cells (unclotted) from a FNA using a needle that can be directly inserted into the tubular container 10, in some embodiments. The cells may then clot (onsite at a patient collection site, for example). A supernatant, e.g., a solution of fixative liquid that may comprise formalin or other suitable fixative material such as Zinc can be introduced into the container 10. Other fixatives may include, but are not limited to, saline, alcohols, acetone, mercury based reagents, and even media (Lennox broth, RPMI, etc.). The container 10 can 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 container for cell processing should be sterilized.

[00120] There may be undesired floating cells in the solution above the clotted cells on the floor 33f of the shaped recess 33 forming the cell bed.

[00121] The lid 40 can be a rigid closed lid that is attached after the fixative is introduced. However, in some embodiments the lid 40 can include a liquid entry port to allow the liquid to enter while the lid remains on. The lid 40 can 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.

[00122] The cell pellet P can be prepared with standard and well known methods with direct access to the base 30 (FIG. 20) and/or with the tubular member attached to the base 30 (FIG. 5B).

[00123] The container 10 with the cell sample can be sent to a cytology laboratory or other suitable research or clinical laboratory. As shown in FIG. 21A and FIG. 21B, the container 10 can be placed in a centrifuge 220 for centrifugation. The centrifuge 220 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 as is known to those of skill in the art. The container 10 can be placed in tandem side by side arrangements as shown in FIGs. 21A and 21B and may include a cap 40 or not. The centrifuge 220 may be configured to process standard 50 mL or 100 mL conical tubes and the container 10 can be placed therein alone or with an adapter. That is, the container 10 may include a sleeve, adapter, or coupler or may have an external integrated size and/or design that allow it to be placed directly into the "bucket" or standard receptacle of the centrifuge. See, e.g., U.S. Pub. No. 2015/0338321, for examples of processing steps, the contents of which are hereby incorporated by reference as if recited in full herein.

[00124] During processing one or more liquids, i.e., a fixative and/or supernatant liquid can be removed by removing the tubular member 20 to expose the drainage aperture 32 allowing the liquid to drain through the at least one drainage aperture.

[00125] In other embodiments, the tubular member 20 can remain attached and the fixative liquid removed by evacuation or decanting. If detached or loosened, the tubular member 20 can be reattached or tightened and another liquid such as a rinse solution can be added to the container 10.

[00126] The fixative liquid can be removed and the rinse added via any suitable technique that leaves the cells and/or cell pellet in the container 10 including aspiration tubing, pipette withdrawal, decanting and the like. The supernatant can 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.

[00127] As before, the rinse solution or other liquids can be removed with the tubular member 20 loosened or off or with the lid 40 remaining on the tubular member 20 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 a respective process.

[00128] In some embodiments, a clot blot formed during the collection/post-collection can be used as a cap for a rinse cycle of the container 10.

[00129] The tube 10 with the rinse can be centrifuged. The rinse can be drained by loosening or removing the tubular container 20 from the base 30 or decanting or otherwise removing or withdrawing the liquid with the tubular container 20 attached to the base, leaving cells on the cell bed 33f, typically in pellet form.

[00130] A matrix material and/or supernatant can be added to the container 10 to form a solid cell block. 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 material can 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 matrixes 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, Am J Clin Pathol, 2000; 1114, 599-606, the contents of which are hereby incorporated by reference as if recited in full herein.

[00131] During one or more points in a cell processing process, the base 30 and body of the tubular member 20 can be detached from each other and/or merely loosened relative to each other thereby removing the lip seal 25s (FIG. 2) formed by the lip 25 over the at least one drainage aperture 32 which unseals and/or exposes the at least one drainage aperture 32. The base 30 can be drained of excess liquid, such as supernatant, via the at least one drainage aperture 32, providing direct access to and/or exposing the solid cell block or soft cell pellet on the cell bed of the shaped recess 33 (FIG. 5D, 18A).

[00132] In some embodiments, merely by turning the base upside down, the solid cell block or cell pellet can be slidably removed from the base 30.

[00133] In some embodiments, the medial bottom segment 30b of the base 30 can be sufficiently flexible to allow a user to push inward against the bottom 30b to deform the bottom and push the formed cell block or the cell pellet out of the base 30.

[00134] In some embodiments, the inner wall segment 35i around the shaped recess 33 of the base 30 can be peelably removed from the solid cell bed.

[00135] The resulting cell pellet (also termed cell disk) can be ready for routine processing. The cell pellet can be sliced or cut for preparing slides for staining or other diagnostic protocols. There may be an increased concentration and/or number of cells in the cell pellet or slices thereof that may promote diagnostic capability over smears alone.

[00136] In some embodiments, different volume tubular members 20 can be provided in a package and a user can select an appropriate one for use at the collection site allowing for increased flexibility corresponding to the specimen type (e.g., urine, blood plasma or serum versus FN A).

[00137] The container 10 can have a defined capacity or volume. The container 10 may 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 tubes 10 can be provided in different volumes/sizes for different applications. [00138] In some embodiments, the container 10 is 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.

[00139] In some embodiments, the containers 10 can be used to process cells for human or veterinary uses. In certain embodiments, the containers 10 can be directed to preparation of cells for pathology review. While it is contemplated that the containers 10 are 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 containers 10.

[00140] 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.

[00141] 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 R A/DNA research or analysis and may include live cells. With the use of appropriate media, the containers 10 can 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.

[00142] In particular embodiments, the cell samples are from endocervical curettages (ECC). 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— pl6— 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, which is also known as (although technically incorrect) immunohistochemistry: pi 6 is a

protein/antigen with the pi 6 antibody in a cell having clinical significance.

[00143] FIG. 17 is a flow chart of exemplary operations that can be used to carry out embodiments of the invention. A container with a tubular member sealably attached to the base is provided. The tubular member comprises a lip that sealably resides over at least one drainage aperture that is above a shaped recess in the base, the shaped recess forming a cell bed, the container comprising a biosample comprising cells that reside in the shaped recess on the cell bed (block 300). The biosample in the tubular container can be centrifuged so that cells from the biosample deposit as a pellet against the cell bed (block 310). A liquid (which can comprise supernatant) can be inserted into the tubular container above the cell bed (block 320).

[00144] A solid cell pellet can be formed holding distributed cells therein on and/or above the floor of the shaped recess (block 330). The tubular member can be loosened from the base to unseal the lip from the at least one drainage aperture in the base (block 340). Liquid (i.e., supernatant) can be drained from the base via at least one drainage aperture above the cell bed in response to the loosening (block 350). The base can be detached from the tubular member (block 360). Direct access to the solid cell pellet can be provided and/or the solid cell pellet can be removed from the base (block 370).

[00145] In some embodiments, merely turning the base over can allow the solid cell pellet to slidably exit ("pour" out of) the base without any other forces applied.

[00146] In some embodiments, the medial bottom of the base can be sufficiently flexible to be able to deform in shape and a user can push against the external bottom of the base, i.e., up or down depending on the orientation of the base, to push the solid cell pellet out.

[00147] In some embodiments, the base 30 can be peelably removed from the solid cell pellet.

[00148] The foregoing is illustrative of embodiments of the present invention and is not to be construed as limiting thereof. Although a few exemplary embodiments of this invention have been described, those skilled in the art will readily appreciate that many modifications are possible in the exemplary embodiments without materially departing from the novel teachings and advantages of this invention. Accordingly, all such modifications are intended to be included within the scope of this invention as defined in the claims. The invention is defined by the following claims, with equivalents of the claims to be included therein.

Claims

THAT WHICH IS CLAIMED:

1. A container comprising:

a tubular member comprising opposing first and second end portions and an open fluid through channel, wherein the second end portion comprises a radially extending lip; and a base releasably and sealably attachable to the tubular member, wherein the base comprises:

a medial portion with a shaped recess having a closed bottom surface; and at least one drainage aperture above the closed bottom surface of the shaped recess,

wherein, when assembled to the tubular member, the radially extending lip sealably covers the at least one drainage aperture.

2. The container of Claim 1, wherein the base further comprises an outer wall and an inner wall, wherein the outer wall comprises threads that attach to mating threads of the tubular member to releasably attach to the tubular member, wherein the inner wall comprises a first wall segment forming an upwardly extending wall of the shaped recess and a second wall segment facing the outer wall, wherein the first inner wall segment is joined to the second inner wall segment by a peak wall segment, and wherein the at least one drainage aperture resides at least partially on the peak wall segment.

3. The container of Claim 1 or 2, wherein the lip extends at least partially about a circumference of the fluid through channel.

4. The container of any of Claims 1-3, wherein the lip extends about an entire circumference of the fluid through channel and has a free inner end that tapers down and inward from an outer wall of the tubular member.

5. The container of any of Claims 1-4, wherein the tubular member has a monolithic molded polymer body with the lip moldably integral therewith.

6. The container of any of Claims 1-5, wherein the second end portion of the tubular member comprises threads that extend at least partially below an inner end of the lip.

7. The container of any of Claims 1-6, wherein the at least one drainage aperture comprises a plurality of circumferentially spaced apart drainage apertures.

8. The container of any of Claims 1-7, wherein the at least one drainage aperture has an elongate radially extending shape, and wherein an end facing the outer wall has a greater diameter than an end facing the shaped recess.

9. The container of any of Claims 1-8, wherein the base further comprises:

an outer wall;

an inner wall; and

an annular cavity between the outer wall and the inner wall,

wherein the outer wall surrounds the inner wall, wherein the inner wall comprises first and second radially spaced apart wall segments joined by a peak wall segment, the first wall segment defining an interior wall of the shaped recess, and wherein the at least one drainage aperture can extend radially at least partially across the peak segment.

10. The container of Claim 9, wherein the second end portion of the tubular member extends a distance into the annular cavity.

11. The container of Claim 1 , wherein the shaped recess has an aspect ratio h/w of between 1 and 1.5.

12. The container of Claim 1, wherein the tubular member and attached base provide a liquid volumetric capacity that is between about 10 mL to about 100 mL.

13. The container of Claim 9, wherein the at least one drainage aperture comprises a plurality of circumferentially spaced apart drainage apertures, wherein the drainage apertures have an elongate radially extending shape.

14. The container of Claim 1, wherein the base has an axially extending centerline, and wherein the bottom of the shaped recess has a radius of curvature "R" measured from the centerline that is between 1-6 mm.

15. The container of any of Claims 1-14, wherein the base comprises a plurality of external "V" shaped partition walls providing a planar bottom surface for the base.

16. The container of any of Claims 1-15, further comprising a cap configured to attach to the first end portion of the tubular member, wherein the tubular member has a length that is greater than that of the base.

17. A method of collecting and processing a biosample, comprising:

(a) providing a container with a tubular member sealably attached to a releasable base, the base comprising at least one drainage aperture above a shaped recess in the base, the shaped recess forming a cell bed, the tubular member comprising a lip that sealably covers the at least one drainage aperture, wherein the container comprises a biosample comprising cells that reside in the shaped recess;

(b) centrifuging the biosample in the tubular container so that cells from the biosample deposit as a pellet against the cell bed;

(c) inserting a liquid material in the tubular container above the cell bed;

(d) forming a solid cell pellet holding distributed cells therein;

(e) loosening the tubular member from the base to unseal the lip from the at least one drainage aperture;

(f) draining liquid from the base via at least one drainage aperture above the cell bed in response to the loosening action;

(g) detaching the base from the tubular member; and then

(h) removing the solid cell pellet from the base.

18. The method of Claim 17, wherein the draining comprises flowing liquid in linear parallel streams for at least an external distance downward from circumferentially spaced apart drainage apertures in the base.

19. The method of Claim 17 or 18, wherein the biosample comprises cells from a fine needle aspirate tissue sample.

20. A method of collecting a biosample, comprising:

providing the container of Claim 1 ; inserting a needle with a fine needle aspirate (FNA) sample comprising cells into the tubular container so that cells reside on the cell bed as the cells of the biosample; and

placing a cap on the container before or after inserting the biosample.

21. A tubular member for a specimen processing container, comprising:

opposing first and second end portions and an open fluid through channel,

wherein the second end portion comprises external threads on a wall thereof and a moldably integral, radially extending internal lip that extends at least partially about a circumference of the fluid through channel.

22. The tubular member of Claim 21, wherein the lip extends about an entire circumference of the fluid through channel and has a free inner end that tapers down and inward from the wall of the tubular member.

23. The tubular member of Claim 21 or 22, wherein the lip has a radial extent in a range of about 3mm-5 mm, measured from an inner surface of the wall of the tubular member.

24. A base for a cell processing container, comprising:

an outer wall and an inner wall spaced apart by an annular cavity, wherein the outer wall comprises internally extending threads, wherein the inner wall comprises a first wall segment forming an upwardly extending wall of the shaped recess and a second wall segment facing the outer wall, wherein the first inner wall segment is joined to the second inner wall segment by a peak wall segment, and wherein the base further comprises a plurality of circumferentially spaced apart drainage apertures residing at least partially across the peak wall segment.

25. The base of Claim 24, wherein the at least one drainage aperture has an elongate radially extending shape, and wherein an end facing the outer wall has a greater diameter than an end facing the shaped recess.

26. The base of Claim 24 or Claim 25, wherein the shaped recess has an aspect ratio h/w of between 1 and 1.5.

27. The base of any of Claims 24-26, wherein the outer wall has a greater height than the inner wall.

28. The base of any of Claims 24-27, wherein the base has an axially extending centerline, and wherein the bottom of the shaped recess has a radius of curvature "R" measured from the centerline that is between 1-6 mm.

29. The base of any of Claims 24-28, wherein the base comprises a plurality of external "V" shaped partition walls, each V-shaped wall extending about one of the drainage apertures, wherein the V-shaped partition walls have a planar external bottom surface providing a planar bottom surface for the base.