Histological tissue sample holding device and method for preparing one or more biopsy tissue sample

Abstract

SECTIONABLE CASSETTE AND EMBEDDING FRAME WITH CONNECTORS AND METHODS FOR PREPARING BIOPSY TISSUE SAMPLES. The present invention relates to a histological tissue sample holder assembly (10), which includes a tissue cassette (12), a frame (14) and a lid (18). The tissue cassette (12) has a recess (11) which includes a body with at least one side wall (16a, 16b, 16c, 16d) and a bottom wall (12b) and is formed of material that can be successfully sectioned in a microtome and is resistant to degradation by solvents and chemicals used to fix, process and stain tissues. The tissue cassette (12) is movably coupled to the frame (14) by a frame-cassette connector (64). The cover (18) is attached to a peripheral portion of the frame (14) by a cover-frame connector (36).The lid (18) and the tissue cassette (12) are able to move from a first position to a second position relative to the frame (14) and in the second position the bottom wall (12b) and at least part of the side wall (16a, 16b, 16c, 16d) extend beyond a bottom edge (14e) of the frame (14) for sectioning in a microtome.

Description

1 / 53 “SAMPLE SUPPORT DEVICE HISTOLOGICAL TISSUE AND METHOD FOR PREPARING One or more biopsy tissue samples. DESCRIPTIVE REPORT Cross-Reference to Related Orders

[0001] This Application claims priority to the Patent Application of U.S. Serial No. 16 / 680601, filed November 12, 2019, the description of which is hereby incorporated by reference in its entirety. Technical Field

[0002] The present invention generally relates to supports for handling and incorporating tissue samples for pathological analysis and, more particularly, to microtome-sectionable supports that can receive one or more tissue samples, and a support frame with a tissue immobilizing cover. Background

[0003] To accurately diagnose various diseases and tissue conditions, medical personnel must remove one or more tissue samples from a patient's body. This process of collecting tissue from the body is known as a biopsy. Once the tissue sample or samples are removed and sent to a pathology laboratory, the tissue will undergo a series of procedures performed by a histotechnician and, finally, a pathologist, to diagnose one or more conditions associated with the tissue. The present invention generally relates to the procedures that are normally performed by the histotechnician to prepare the tissue sample or samples on slides. Petition 870250039567, dated 05 / 15 / 2025, page 63 / 118 2 / 53 which can be analyzed under a microscope by a pathologist.

[0004] Although the singular term “sample” is used throughout this report, it should be understood that this term also encompasses plural “samples.” Once a tissue sample is removed from a patient’s body, it is typically placed in a sample container containing a tissue fixative solution, and then the container is transported to a pathology laboratory. The tissue will undergo a process known as “grossing-in” in the pathology laboratory, during which a histotechnician will retrieve the tissue sample from the container, typically cut the tissue into sizes appropriate for tissue processing, place individual samples into small plastic tissue cassettes of appropriate size, and assign tracking numbers to each cassette. The assignment of tracking numbers is usually done by printing the tracking number on the cassette or on a label that is then applied to the cassette.These tracking numbers are then recorded in a tracking system used in the laboratory. For the smallest tissue samples, which may be just scrapings, the cassette includes fine mesh openings on the sides and bottom. In other situations involving very small tissue samples, the samples are placed in a bag that resembles a tea bag, which prevents the smallest tissue samples from escaping. Larger tissue samples are placed in cassettes with slightly larger notched openings that are, however, smaller than the tissue sample inside the cassette.

[0005] The cassettes are then placed in a perforated stainless steel basket and passed through a tissue processing machine, often overnight. This machine uses a combination of vacuum, heat, and liquid reagents or chemicals to remove interstitial fluids from within the tissue. Once the fluids have been removed from the tissue samples, the processing machine immerses the tissue samples in a bath of a Petition 870250039567, dated 05 / 15 / 2025, page 64 / 118 3 / 53 hardenable material, such as molten paraffin (i.e., a form of wax), so that the interstices in the tissue are replaced by paraffin. The histotechnician then removes the basket from the machine and removes the individual tissue cassettes. In a conventional procedure practiced for many years, the histotechnician individually removes the tissue sample from each cassette. The histotechnician must carefully orient the tissue sample, based on the tissue type, into a stainless steel-based mold that is approximately the size of the tissue cassette and partially loaded with molten paraffin. The tissue sample must be held manually, usually using forceps, against the bottom of the mold. Failure to do so may compromise the ability to make adequate slices of the tissue sample later on a microtome.The molten paraffin is then rapidly cooled on a chilled plate, which may be an electric thermal chiller (TEC), to partially solidify the paraffin, thus keeping the tissue sample in the proper orientation against the bottom of the mold.

[0006] The cassette is then placed on top of the base mold, and an embedding material, which is also typically paraffin wax, is poured through the top of the open cassette into the base mold. The cassette changes its function at this point in the procedure, from a tissue retention component to a fixation-type device for mounting in the microtome and producing shavings or slices of the solidified paraffin block (containing the tissue sample) in the microtome. The base mold is cooled until all the molten paraffin has hardened, and the histotechnician removes the stainless steel base mold from the paraffin block and embedded tissue. The tissue sample is then embedded within a rectangular block of hard paraffin with a plastic tissue cassette on the opposite side. As mentioned, the cassette can then be used as a support or fixation in the microtome mandrel.Just as with the tissue processing machine, the embedding process is carried out in batches, during which a histotechnician can... Petition 870250039567, dated 05 / 15 / 2025, page 65 / 118 4 / 53 process an average of approximately 40 to 60 cassettes per hour in embedded tissue blocks.

[0007] The hardened paraffin blocks containing the embedded tissue samples are therefore ready to be cut into extremely thin sections for placement on a microscope slide. The histotechnician mounts the embedded tissue block on a mandrel in the microtome sized to receive the side of the block containing the embedded plastic cassette. The histotechnician can then begin slicing the paraffin block that has the embedded tissue sample opposite the surface of the plastic cassette. This produces a strip of individual slices of the embedded tissue in the hardened paraffin. The action of the microtome causes the individual slices to fuse together when done correctly, and subsequently these very thin strips of slices are immersed in a water bath and a glass slide is carefully placed under the slice. Each slice, with the finely sectioned tissue sample embedded in it, is then adhered to the top of a microscope slide.When the histotechnician has enough slides of the tissue sample, the slides are placed in an automated staining machine. The staining machine goes through a series of infiltration steps to stain the different tissues and cells of the slide with different colors. This helps the pathologist identify different structures and makes it easier to find any abnormalities in the tissue. After the staining procedure is complete, the slides are covered by a slide cover and prepared for the pathologist to place under a microscope for analysis.

[0008] Based on the summary of the procedure provided above, it will be observed that the conventional handling and processing of tissue samples is a very laborious process involving several manual steps performed by a histotechnician. Thus, repetitive strain injuries, such as carpal tunnel syndrome, are prevalent. This is especially true with the tissue sample embedding process. These multiple manual operations and the Petition 870250039567, dated 05 / 15 / 2025, page 66 / 118 5 / 53 Repetitive handling of tissues increases the likelihood of human error and, moreover, requires highly trained and qualified histotechnicians to ensure that tissue samples, essentially adhered to slides for analysis by the pathologist, are in ideal conditions and orientation to obtain diagnostic accuracy.

[0009] U.S. Patents Nos. 5,817,032 (the '032 patent), 7,156,814, 7,179,424, 7,722,810, 7,776,274 and 8,383,067, and thePublication of U.S. Patent Application No. 2018 / 0156701 describes several improvements to this area of ​​technology, including novel ways of containing tissue samples during grossing-in, embedding, and microtome or slicing procedures. The descriptions of patents '032, 7,156,814, 7,179,424, 7,722,810, 7,776,274, 8,383,067, and 2018 / 0156701 are hereby fully incorporated herein by reference. For example, patent '032 refers to a tissue trapping and support device, which may be a cassette and which may be successfully sectioned using a microtome. When such a cassette is used, the tissue sample is immobilized within the cassette and subjected to the process of tissue fluid replacement with paraffin. Next, both the tissue sample and the cassette are sliced ​​simultaneously for later mounting on microscope slides.Since the tissue sample is never removed from the cassette from the moment it is processed in the tissue processing machine until the moment it is cut or sliced ​​with the microtome, a significant amount of handling time is saved. Furthermore, the chance of human error or tissue loss is significantly reduced due to the elimination of separate tissue handling steps. Patent '032 and the other patent properties incorporated above generally also describe other improvements that help automate the overall process and, in conjunction with the new tissue holders (e.g., cassettes), can further reduce handling steps throughout the procedure and make the procedure more reliable. Petition 870250039567, dated 05 / 15 / 2025, page 67 / 118 6 / 53

[00010] There are several disadvantages to current procedures and limitations to innovation. For example, improvements in the external shape of the cassette and frames are restricted by existing limitations in histopathology laboratory equipment, such as tissue processing retorts and “entry devices” for tissue processors, embedding stations, printers, and microtomes. Many of these processes are integrated into systems and machines for automation of steps and robotic manipulation, further limiting the potential for innovation. In addition, material costs have increased in recent years, especially for fluoropolymer sectionable plastics (FEP / PFA) useful in sectionable cassettes. Each cassette is essentially consumed by the sectioning procedure, which increases the cost of the pathology procedure.Furthermore, since the sectionable FEP / PFA material is not rigid, it can be used for some components (e.g., the cassette), while other components (e.g., frames) can be constructed from harder or more rigid materials.

[00011] Despite the various advances made in this field, there is a need for further improvements related to inclusion cassettes and frames, particularly for cassette and frame assemblies including some components made of relatively soft materials (e.g., sectionable plastics) that are coupled to other components made of more rigid materials. SUMMARY

[00012] According to one embodiment, a histological tissue sample holder device includes a tissue cassette with a recess including at least one side wall and one bottom wall. The tissue cassette is formed of a first material that can be successfully sectioned in a microtome and is resistant to degradation by solvents and chemicals used for fixation, processing, and staining. Petition 870250039567, dated 05 / 15 / 2025, pages 68 / 118 7 / 53 the tissue. The device further includes a frame including a lower edge, the frame being formed of a second material different from the first material and more rigid than the first material. The tissue cassette is attached to the frame by a frame-cassette connector, including a first retention structure formed integrally with the frame that extends, at least partially, through a second retention structure formed integrally with the cassette. The device further includes a lid attached to the frame. The lid and the tissue cassette are capable of moving from a first position to a second position relative to the frame. In the second position, the lower wall and at least part of the side wall extend downwards beyond the lower edge of the frame for sectioning in the microtome. The frame can be detached from the cassette by separating the frame-cassette connector.

[00013] In additional or alternative aspects, the first retaining structure may include a pin integrally formed with the frame. The second retaining structure may include a flange integrally formed with the cassette. Moving the cover and fabric cassette from the first position to the second position may break the flange integrally formed with the cassette. The flange integrally formed with the cassette may include a tension elevator arranged to promote the breakage of the flange integrally formed with the cassette when the cover and fabric cassette are moved from the first position to the second position. The frame may include a plurality of outer walls that generally extend upwards from the lower edge. The pin integrally formed with the frame may be arranged in a tab that generally extends laterally into one of the plurality of outer walls.The pin, integral with the frame, can generally extend upwards from the flange. The pin, integral with the frame, can generally extend downwards from the flange. The flange extending from one of the plurality of outer walls can be coupled in a way. Petition 870250039567, dated 05 / 15 / 2025, pp. 69 / 118 8 / 53 articulated to one of the plurality of external walls. The pin formed integrally with the frame may have a generally circular cross-section. The pin formed integrally with the frame may include a base, a tip with a tip width, and a shaft with a shaft width and extending from the base to the tip, and the tip width may be greater than the shaft width.

[00014] In additional or alternative aspects, the cover may be coupled to the frame by a cover-frame connector comprising a third retaining structure integrally formed with a peripheral part of the frame extending, at least partially, through a fourth retaining structure integrally formed with the cover. The cover may be decoupled from the peripheral part of the frame by separating the cover-frame connector. The third retaining structure may include a pin integrally formed with the peripheral part of the frame. The fourth retaining structure may include a flange integrally formed with the cover. Moving the cover and the fabric cassette from the first position to the second position may break the flange integrally formed with the cover. The cover may be formed from the first material. The peripheral part of the frame may include a plurality of peripheral walls surrounding the cover.The pin, integral with the peripheral part, may be disposed in a tab that generally extends laterally into one of the plurality of peripheral walls. When the cover is in a closed configuration, the pin, integral with the peripheral part, may generally extend downwards from the tab. When the cover is in a closed configuration, the pin, integral with the peripheral part, may generally extend upwards from the tab. The tab extending from one of the plurality of peripheral walls may be articulated to one of the plurality of peripheral walls. The pin, integral with the peripheral part, may generally have a circular cross-section. Petition 870250039567, dated 05 / 15 / 2025, pp. 70 / 118 9 / 53

[00015] In additional or alternative aspects, the fabric cassette may include a cassette closing element and the lid may include a lid closing element, the cassette closing element and the lid closing element being configured, when engaged, to secure the lid to the fabric cassette. One of the cassette closing element and the lid closing element may include a first connector disposed on a first extension arm, and the other of the cassette closing element and the lid closing element may include a second connector, the first connector engaging with the second connector to secure the lid to the fabric cassette. The lid closing element may include the first connector disposed on the first extension arm and the cassette closing element may include the second connector.The lid closing element may include a third connector arranged on a second extension arm, the first extension arm and the second extension arm generally projecting downward from the lid when the lid is in a closed configuration on the cassette, the first extension arm and the second extension arm arranged on the lid in an opposite arrangement and spaced apart so that the first connector faces away from the third connector. The cassette closing element may include a fourth connector arranged to engage with the third connector, the second connector and the fourth connector generally being oriented laterally.

[00016] In additional or alternative aspects, the lid closing element may include a third connector arranged on a second extension arm, the first extension arm and the second extension arm generally projecting downwards from the lid when the lid is in a closed configuration on the cassette, the first extension arm and the second extension arm arranged on the lid in an opposite arrangement, so that the first connector faces the third connector. The second connector Petition 870250039567, dated 05 / 15 / 2025, pp. 71 / 118 10 / 53 can generally be oriented laterally and can be arranged to engage with the first connector and the third connector. The frame may include a plurality of outer walls generally extending upwards from the bottom edge and a peripheral part coupled to one of the plurality of outer walls by a hinge. The fabric cassette may be coupled to one of the plurality of outer walls and the cover may be coupled to the peripheral part. At least one of the plurality of outer walls may include a frame closing element, and the peripheral part may include a peripheral part closing element, the frame closing element and the peripheral part closing element being configured, when engaged in a closed configuration, to secure the peripheral part to the plurality of outer walls.One of the frame closing element and the perimeter closing element may include a latch disposed on an extension arm, and the other of the frame closing element and the perimeter closing element may include a flange, the latch engaging the flange to secure the perimeter to the plurality of external walls in the closed configuration. The perimeter closing element may include a latch disposed on the extension arm, and the frame closing element may include the flange. The extension arm may generally project downwards from the perimeter in the closed configuration, and the flange may generally be oriented laterally.

[00017] In additional or alternative aspects, the cover may include a plate configured to be received within the recess of the tissue cassette, the plate being mounted on a peripheral part of the cover by a plurality of protrusion members arranged to incline the plate towards the lower wall of the recess when the cover is in a closed configuration. The protrusion members may be arranged in at least one of a generally helical arrangement and a generally perpendicular arrangement between the plate and the Petition 870250039567, dated 05 / 15 / 2025, pp. 72 / 118 11 / 53 peripheral part of the lid.

[00018] According to another embodiment, a histological tissue sample holder device includes a tissue cassette with a recess including at least one side wall and one bottom wall. The tissue cassette is formed of a first material that can be successfully sectioned on a microtome and is resistant to degradation by solvents and chemicals used to fix, process, and stain the tissue. The device further includes a frame that includes a bottom edge. The tissue cassette is movably attached to the frame. The device further includes a lid attached to the frame. The lid includes a plate configured to be received within the recess of the tissue cassette. The plate is mounted on a peripheral part of the lid by a plurality of protrusion members arranged to tilt the plate toward the bottom wall of the recess when the lid is in a closed configuration.The lid and tissue cassette are capable of moving from a first position to a second position relative to the frame. In the second position, the bottom wall and at least part of the side wall extend beyond the bottom edge of the frame for sectioning in the microtome.

[00019] In additional or alternative aspects, the protrusion members may be arranged in a generally helical arrangement between the plate and the peripheral part of the lid. The protrusion members may be arranged in a generally perpendicular arrangement between the plate and the peripheral part of the lid. The plate may include a plurality of teeth extending towards the lower wall of the recess when the lid is in the closed configuration. The lid may be formed from the first material. The frame may be formed from a second material different from the first material and more rigid than the first material, the fabric cassette being coupled to the frame by a frame-cassette connector comprising a first retaining structure formed integrally with the frame extending through a second retaining structure formed integrally Petition 870250039567, dated 05 / 15 / 2025, pp. 73 / 118 12 / 53 with the cassette. The first retaining structure may include a pin integrally formed with the frame, and the second retaining structure may include a flange integrally formed with the cassette. The cover may be coupled to the frame by a cover-frame connector comprising a first retaining structure integrally formed with a peripheral part of the frame that extends, at least partially, through a second retaining structure integrally formed with the cover. The cover may be decoupled from the peripheral part of the frame by separating the cover-frame connector. The first retaining structure may include a pin integrally formed with the peripheral part of the frame. The second retaining structure may include a flange integrally formed with the cover.

[00020] The invention further provides a method for manufacturing an apparatus for containing a histological tissue sample while sectioning the tissue sample in a microtome. The method includes molding a tissue cassette having a recess including at least one side wall and one bottom wall, the tissue cassette being formed of a first material that can be successfully sectioned in a microtome and that is resistant to degradation by solvents and chemicals used to fix, process and stain the tissue. The method further includes molding a frame including a bottom edge, the frame being formed of a second material different from the first material and more rigid than the first material.The method further includes attaching the fabric cassette to the frame by assembling a frame-cassette connector comprising a first retention structure formed integrally with the frame extending, at least partially, through a second retention structure formed integrally with the cassette. The cassette is capable of being detached from the frame by separating the frame-cassette connector.

[00021] In additional or alternative aspects, the first Petition 870250039567, dated 05 / 15 / 2025, pp. 74 / 118 13 / 53 The retaining structure may include a pin integrally formed with the frame. The second retaining structure may include a flange integrally formed with the cassette. The assembly of the frame-cassette connector may include forming a mushroom-head shaped tip on the pin integrally formed with the frame to secure the flange integrally formed with the cassette onto the pin integrally formed with the frame. Forming the mushroom-head shaped tip on the pin may include deforming the pin to form the mushroom-head shaped tip. Deforming the pin to form the mushroom-head shaped tip may include using a mushroom-head shaped tip forming tool when the pin is at a temperature above ambient temperature and below the melting temperature of the second material.The assembly of the frame-cassette connector may include at least one of (1) assembly of separately molded components, (2) co-molding of the frame and cassette, and (3) insert molding of the frame and cassette. The method may further include molding a cap; and attaching the cap to the frame by assembling a cap-frame connector comprising a third retaining structure integrally formed with a peripheral part of the frame extending through a fourth retaining structure integrally formed with the cap. The cap may be decoupled from the peripheral part of the frame by separating the cap-frame connector. The third retaining structure may include a pin integrally formed with the peripheral part of the frame. The fourth retaining structure may include a flange integrally formed with the cap.The assembly of the cap-frame connector may include at least one of (1) assembly of separately molded components, (2) co-molding of the cap and frame, and (3) insert molding of the cap and frame.

[00022] The invention further provides a method for preparing one or more biopsy tissue samples for histological examination. The method includes positioning a tissue sample in a cassette of Petition 870250039567, dated 05 / 15 / 2025, pages 75 / 118 14 / 53 tissue having a recess including at least one side wall and one bottom wall, the tissue cassette formed of a first material that can be successfully sectioned in a microtome and is resistant to degradation by solvents and chemicals used to fix, process and stain the tissue, the tissue cassette being disposed in a frame including a bottom edge, the tissue cassette being coupled to the frame by a frame-cassette connector that includes a first retaining structure formed integrally with the frame extending through a second retaining structure formed integrally with the cassette. The method further includes closing a peripheral part of the frame, the peripheral part of the frame including a cassette cover disposed thereon. The method further includes moving the cover and the tissue cassette from a first position to a second position relative to the frame, including breaking the frame-cassette connector.In the second position, the lower wall and at least part of the side wall extend downwards beyond the lower edge of the frame for sectioning in the microtome.

[00023] In additional or alternative aspects, the first retention structure may include a pin formed integrally with the frame. The second retention structure may include a flange formed integrally with the cassette. The handling operation may include breaking a lid-frame connector including a third retention structure formed integrally with the peripheral part of the frame extending through a fourth retention structure formed integrally with the lid. The third retention structure may include a pin formed integrally with the peripheral part of the frame. The fourth retention structure may include a flange formed integrally with the lid. The frame may be formed from a second material different from the first material and more rigid than the first material; and the lid may be formed from the first material. The closing operation may include securing the tissue sample in the recess using a plate mounted on the lid by Petition 870250039567, dated 05 / 15 / 2025, pp. 76 / 118 15 / 53 a plurality of propensity members arranged to tilt the plate towards the lower wall of the recess.

[00024] Several additional features and advantages of the invention will become more apparent to those skilled in the art upon review of the following detailed description of the illustrative embodiments taken in combination with the accompanying drawings. Brief Description of the Drawings

[00025] FIG. 1 is a perspective view of an assembly / set according to a particular embodiment.

[00026] FIG. 1A is an exploded perspective view of the assembly in FIG. 1.

[00027] FIG. 2 is a top view of the assembly in FIG. 1, showing the fabric cassette and frame in the open position, ready to receive the fabric in the fabric cassette.

[00028] FIG. 3A is a cross-sectional view of the assembly of FIG. 1, taken generally along line 3-3 of FIG. 2, showing the tissue cassette and frame in a partially closed position with the tissue in the tissue cassette.

[00029] FIG. 3B is a cross-sectional view similar to FIG. 3A, but showing the perimeter of the frame and the cover in the closed position.

[00030] FIG. 3C is a cross-sectional view similar to FIG. 3B, but showing the assembly in the closed position. A portion is further separated to show details of the connector.

[00031] FIG. 3D is a cross-section similar to FIG. 3C, but showing the assembly in a staggered position or in the second position, where the lid is separated from the peripheral part of the frame, and a part of the tissue cassette is ready to be incorporated and then sectioned in a microtome.

[00032] FIG. 4A is a cross-sectional view of the assembly. Petition 870250039567, dated 05 / 15 / 2025, pp. 77 / 118 16 / 53 of FIG.1, usually taken along line 4A-4A of FIG. 2 and in the open position.

[00033] FIG. 4B is a cross-sectional view similar to FIG. 4A, but showing the perimeter of the frame and the cover in the closed position.

[00034] FIG. 4C is a cross-sectional view similar to FIG. 4B, but showing the assembly in the staggered position or in the second position, where the cover is separated from the peripheral part of the frame.

[00035] FIGS. 5A-5D are detailed cross-sectional views of the assembly in FIG. 1, taken generally along line 5-5 in FIG. 2, showing the steps of an assembly process.

[00036] FIG. 5E is a detailed cross-sectional view of the assembly in FIG. 1, taken generally along line 5E-5E of FIG. 2 with the cover in the closed position.

[00037] FIGS. 6A-6D are a cross-sectional view of an alternative pin / flange connector showing the steps of an assembly process.

[00038] FIGS. 7A-7C are a cross-section of another alternative pin connector showing the steps of an assembly process.

[00039] FIGS. 7D-7F are a cross-sectional view of another alternative pin connector showing the steps of an assembly process.

[00040] FIG. 8 is a perspective view of an alternative assembly according to another embodiment.

[00041] FIG. 9 is a top view of the assembly in FIG. 8, showing the fabric cassette and frame in the open position, ready to receive the fabric in the fabric cassette.

[00042] FIG. 10A is a cross-sectional perspective view of the assembly in FIG. 8, generally taken along line 10A-10A in FIG. 9 and in the open position. Petition 870250039567, dated 05 / 15 / 2025, pages 78 / 118 17 / 53

[00043] FIG. 10B is a cross-sectional perspective view similar to FIG. 10A, but showing the perimeter of the frame and the lid in the closed position.

[00044] FIG. 10C is a cross-sectional view similar to FIG. 10B, but showing the assembly in the stepped position or in the second position, where the cover is separated from the peripheral part of the frame.

[00045] FIG. 11 is a perspective view of an alternative assembly according to another embodiment.

[00046] FIG. 12 is a top view of the assembly in FIG. 11, showing the fabric cassette and frame in the open position ready to receive the fabric in the fabric cassette.

[00047] FIG. 13A is a cross-sectional view of the assembly of FIG. 11, taken generally along line 13A-13A of FIG. 12, showing the fabric cassette and frame in a partially closed position.

[00048] FIG. 13B is a cross-sectional view of the assembly of FIG. 11, usually taken along line 13B-13B of FIG. 12, but showing the assembly in a staggered position or in the second position, where the lid is separated from the peripheral part of the frame and a portion of the tissue cassette is ready to be incorporated and then sectioned in a microtome.

[00049] FIG. 14A is a cross-sectional perspective view of the assembly in FIG. 11, taken generally along line 14A-14A of FIG. 12 and in the open position.

[00050] FIG. 14B is a cross-sectional perspective view similar to FIG. 14A, but showing the perimeter of the frame and the lid in the closed position.

[00051] FIG. 14C is a cross-sectional perspective view similar to FIG. 14B, but showing the assembly in the staggered position or in the second position, where the cover is separated from the peripheral part of the frame. Petition 870250039567, dated 05 / 15 / 2025, pp. 79 / 118 18 / 53

[00052] FIG. 15 is an exploded perspective view of an alternative assembly according to another embodiment.

[00053] FIG. 16A is a detailed exploded perspective view of the lid and perimeter frame of FIG. 15.

[00054] FIG. 16B is a detailed perspective view of a cover-frame connector of the assembly in FIG. 15. Detailed Description

[00055] Referring first to FIGS. 1, 1A and 2, an assembly 10, constructed according to an illustrative embodiment of the invention, is shown in the open position. The assembly 10 includes a tissue sample cassette 12 carried within and separably coupled to a frame 14, which includes a peripheral part 16. A lid 18 is separably coupled to the peripheral part 16. Although the cassette 12 has a rectangular configuration, it has been recognized that the cassette 12 may have alternative configurations. For example, a cassette may have a circular configuration. The peripheral part 16 generally includes an interior 11 defined between the surrounding (peripheral) side walls 16a, 16b, 16c, 16d, and the cover 18 is sized and configured to fit within the interior and is separably coupled to at least one of the surrounding walls 16a, 16b, 16c, 16d.The frame 14 generally includes a defined interior between the surrounding outer walls 14a, 14b, 14c, 14d and a lower edge 14e, and the cassette 12 is sized and configured to move within the interior between at least the first and second positions, as generally discussed in the patent properties incorporated above and for the same purposes. The first position is shown in FIGS. 3B and 3C, while the second, “stepped” position is shown in FIG. 3D. In the second position, the lower part of the cassette 12 is exposed below the lower edge 14e of the frame 14, to allow the cassette 12 and an embedded tissue sample 44 to be sectioned in a microtome. Petition 870250039567, dated 05 / 15 / 2025, pages 80 / 118 19 / 53 while frame 14 is held in the microtome chuck.

[00056] The connection of the fabric cassette 12 to the frame 14 can be carried out in many different ways, such as any of the ways described in the patent properties incorporated above. Alternatively, the cassette 12 can be coupled to the frame 14 in other novel ways, such as those described below. In the illustrative embodiment of FIG. 1, the cassette 12 is initially separably coupled to the frame 14 via frame-cassette connectors 64 that couple the surrounding walls 14a, 14b, 14c, 14d to the cassette 12.

[00057] With reference to FIGS. 4A-4C and 5D, each frame-cassette connector 64 includes a retaining structure (e.g., first retaining structure), such as a pin 66, integrally formed with the frame 14 and extending at least partially through a retaining structure (e.g., second retaining structure), such as a flange 68 integrally formed with the cassette 12, such as on a side wall 12a of the cassette 12.Each pin 66 generally extends upward from a flange 70, which generally extends laterally into one of the outer walls 14a, 14b, 14c, 14d. The pin 66 includes a base 72 disposed on the flange 70, a tip 74 on an opposite surface of the flange 68, and a shaft 76 extending from the base 72 to the tip 74, through the flange 68. The tip 74 may have a tip width 78 that is greater than a shaft width 80. Generally, because the tip 74 and the flange 70 are wider than the shaft 76, and because the flange 68 extends into the recessed area or recess between the tip and the flange, the flange is retained in the pin 66 by the tip and the flange. Consequently, until the cassette-frame connector 64 is separated (e.g., broken) during the scaling operation, as discussed in more detail below, the flange 68 is attached to the pin 66. The pin 66 may be formed as a straight circular cylinder or a truncated cone having a generally circular cross-section.In this illustrative example, the first retaining structure is a pin 66, but it will be. Petition 870250039567, dated 05 / 15 / 2025, pages 81 / 118 20 / 53 It is understood that in other embodiments the first retaining structure may assume other forms, such as any shape or structure configured to extend at least partially through a corresponding second retaining structure (for example, the flange 68 formed integrally with the cassette 12) to secure or couple two or more components. Such first retaining structures may include, for example and without limitation, a tongue, a bar, a shaft, a rod, a stick, etc., and may have any cross-sectional shape, such as generally circular, generally oval, or generally polygonal (for example, generally square, pentagonal, hexagonal, etc.). Furthermore, a first retaining structure may be generally straight or may include one or more curves, bends, or angles.

[00058] With reference to FIGS. 4B (first position) and 4C (second position), the frame-cassette connectors 64 of this illustrative embodiment are frangible and are configured to break when the cassette 12 is moved from the first position to the second position. In the illustrative embodiment, the pin 66 pulled from the flange 68 breaks the flange 68 during this movement. In other embodiments with different relative sizes or material choices for the pin 66 and flange 68, the pin 66 or the tab 70 may break before the pin 66 is pulled from the flange 68. In other embodiments, various connectors, such as frame-cassette connectors and cap-frame connectors (discussed below), may be configured to separate without breaking. For example, some connectors may be configured to accommodate sufficient elastic deformation to allow a pin to be removed from a flange without rupture of the pin, tab, or flange.Returning to the illustrative example, as can be seen by comparing the orientation of tabs 70, FIGS. 4B and 4C, each tab 70 can be articulated to its respective outer wall 14a, 14b, 14c, 14d, so that, during the movement from the first position to the second position, tab 70 articulates downwards. Petition 870250039567, dated 05 / 15 / 2025, page 82 / 118 21 / 53 which can promote a predictable and consistent separation (e.g., breakage) of the frame-cassette connector 64 (e.g., pins 66 pulled from flanges 68).

[00059] The connection of the cover 18 to the peripheral part 16 of the frame 14 can be carried out in many different ways, as well as any of the ways described in the patent properties incorporated above. Alternatively, the cover 18 can be coupled to the peripheral part 16 of the frame 14 in other novel ways, such as those described below. In the illustrative embodiment of FIG. 1, the cover 18 is initially coupled in a separable manner to the peripheral part 16 of the frame 14 by means of cover-frame connectors 36 that couple the surrounding walls 16a, 16b, 16c, 16d to the cover 18.

[00060] The cap-frame connectors 36 can generally be similar in structure and operation to the frame-cassette connectors 64 described above. With reference to FIGS. 4C and 5E, each cap-frame connector 36 includes a retaining structure (e.g., first retaining structure), such as a pin 82, integrally formed with the peripheral part 16 of the frame 14 and extending at least partially through a retaining structure (e.g., second retaining structure), such as a flange 84 integrally formed with the cap 18. Each pin 82 generally extends downwards from a tab 86, which generally extends laterally into one of the surrounding walls 16a, 16b, 16c, 16d. Pin 82 includes a base 88 (see FIG. 5E) disposed in the tab 86, a tip 90 on an opposite surface of the flange 84, and a shaft 92 extending from the base 88 to the tip 90 through the flange 84.The 90 tip can have a tip width of 94, which is greater than a shaft width of 96. The 82 pin can be formed as a straight circular cylinder or a truncated cone with a generally circular cross-section. In this illustrative embodiment, the first retaining structure is an 82 pin; however, it should be understood that in other embodiments the first retaining structure may assume other forms. Petition 870250039567, dated 05 / 15 / 2025, page 83 / 118 22 / 53 forms, such as any shape or structure configured to extend at least partially through a corresponding second retaining structure (e.g., flange 84 formed integrally with cassette 18) to secure or couple two or more components. Such first retaining structures may include, for example and without limitation, a tongue, a bar, a shaft, a rod, a stick, etc., and may have any cross-sectional shape, such as generally circular, generally oval, or generally polygonal (e.g., generally square, pentagonal, hexagonal, etc.). Furthermore, a first retaining structure may be generally straight or may include one or more curves, bends, or angles.

[00061] With reference to FIGS. 4B (first position) and 4C (second position), the cover-frame connectors 36 of this illustrative embodiment are frangible and are configured to break when the cover 18 is moved from the first position to the second position. In the illustrative embodiment, pin 82 pulled from flange 84 breaks flange 84 during this movement. In other embodiments with different relative sizes or material choices for pin 82 and flange 84, pin 82 or tab 86 may break before pin 82 is pulled from flange 84. As can be seen by comparing the orientation of tabs 86, FIGS. 4B and 4C, each tab 86 can be articulated to its respective surrounding wall 16a, 16b, 16c, 16d, so that, during movement from the first position to the second position, the tab 86 articulates downwards, which can promote a predictable and consistent separation (e.g., rupture) of the cover-frame connector 36 (e.g., pins 82 pulled from the flanges 84).

[00062] Now with reference to FIGS. 2, 3A and 3B, the connections between the frame 14 and the peripheral part 16 are described in more detail. The peripheral part 16 is coupled to the wall 14a of the frame 14 by a pair of hinges 22a, 22b, which are optionally frangible. The part Petition 870250039567, dated 05 / 15 / 2025, pages 84 / 118 23 / 53 peripheral part 16 engages under pressure in the closed position (FIG. 3B) through the engagement of the peripheral part's closing elements, which may include connectors such as latches 24, 26, with frame closing elements, which may include connectors such as flanges 30, 32. The latch 24, which may be hook-shaped, is positioned on the outer wall 16a of the peripheral part 16 and engages with the flange 30 of the wall 14a of the frame 14 in the closed position. The latches 26 are positioned on the wall 16c of the peripheral part 16 and engage (e.g., press fit) with a flange 32 of the wall 14c of the frame 14 in the closed position.

[00063] Now with reference to FIGS. 1, 3A and 3B, the connections between the cover 18 and the cassette 12 are described in more detail. The cover 18 is press-fitted into the closed position (FIG. 3B) by means of the engagement of cover locking elements, which may include connectors such as flanges 58, with cassette locking elements, which may include connectors such as latches 60. In this embodiment, the cover 18 includes a pair of opposing flanges 58 arranged to press-fit with two opposing pairs of latches 60 in the closed position. In the closed position, with the flanges 58 of the cover 18 engaged with the latches 60 of the cassette 12, the cover 18 and the cassette 12 are coupled and move between the first position (FIG. 3C) and the second position or stepped position (FIG. 3D) as a single unit.

[00064] The cassette 12 and the cover 18 are dimensioned and configured to move within the frame 14 between at least the first and second positions, as best shown in FIGS. 3C and 3D. With reference to FIGS. 2, 4B and 4C, the cover 18 includes a cover retaining flange 38 at each of its four corners. The retaining flanges 38 are configured to engage with cassette positioning elements 40 of the frame 14, which are formed as part of the inner corners of the four corners of the frame 14. In the illustrative embodiment, each retaining flange 38 engages with a respective cassette positioning element 40. The elements Petition 870250039567, dated 05 / 15 / 2025, page 85 / 118 24 / 53 of cassette positioning 40 are flexible and hollow, so that as the retaining flanges 38 pass through the cassette positioning elements 40 (e.g., downwards), the retaining flanges 38 deform the cassette positioning elements 40 and finally “snap” under the cassette positioning elements 40, as shown, for example, in FIG. 4C.

[00065] As shown in FIGS. 2 and 4A-4C, each corner of the frame 14 includes a diagonal stop 46 located below and away from the downward-sloping part of the respective cassette positioning element 40. When the cassette 12 reaches the second position, the stops 46 prevent the downward movement of the retaining flanges 38 of the lid 18. Thus, in the second position, the retaining flanges 38 of the lid 18 are vertically fixed between the cassette positioning elements 40 (at the top) and the stops 46 (at the bottom). As the lid 18 and the cassette 12 are coupled together by flanges 58 and latches 60, this keeps the cassette 12 and the lid 18 in the second position and ready for embedding and subsequent sectioning in the microtome, as described in the patent properties incorporated above.

[00066] As shown in FIG. 3A, one or more tissue samples 44 can be placed in the cassette 12, which defines a recess or internal area 11 surrounded by at least one side wall 12a and including a bottom wall 12b. Although a generally rectangular recess 11 is shown (see FIG. 2), it will be noted that any other shape, such as any polygon (e.g., square) or any rounded shape (e.g., oval or circular) or shapes with depression or alignment features for the tissue sample 44, can be used in its place.

[00067] This illustrative embodiment also includes a resilient structure carried on the underside of the cap 18. The resilient structure is in the form of curved elastic fingers 42 for the purpose of allowing flexible engagement between the distal ends 42a of the resilient fingers. Petition 870250039567, dated 05 / 15 / 2025, pages 86 / 118 25 / 53 and one or more tissue samples 44 in cassette 12. The resilient fingers 42 form a compliant structure that maintains the tissue 44 in the desired orientation and position without creating an artifact impression on the tissue sample 44 during processing. With reference to FIG. 4A, in this embodiment, the distal ends 42a of resilient fingers include teeth 42b, which can generally extend downwards when the lid 18 is in the closed position. The teeth 42b can provide additional security against unwanted movement of the tissue sample 44.

[00068] It will be observed that different materials and configurations of resilient finger 42 can be chosen based on, for example, the type of tissue to be processed and analyzed. For example, small mucosal tissue samples can be successfully held and processed using some arrangements of resilient fingers 42, while other tissue types, such as adipose tissue, may be better served by another material or configuration of resilient finger 42. As another example, larger tissue samples may require a containment structure that works well over a large surface area. Furthermore, resilient fingers 42 may have specific tissue orientation or containment alignment features to facilitate the orientation of very specific types of tissue samples.Generally, resilient fingers can be arranged on the lid in a uniform or non-uniform arrangement or orientation, can be shaped (e.g., shape, thickness, or length), or can be otherwise modified (e.g., similar to the configurations described above in the incorporated patent properties) as desired to facilitate the acceptance and containment of tissue samples of various types, sizes, or thicknesses.

[00069] The resilient fingers 42 allow the infiltration of solvents and chemicals used to fix, process and stain the tissue and of the embedding material used for tissue embedding, while the tissue is contained by the resilient fingers 42. The resilient fingers 42 are Petition 870250039567, dated 05 / 15 / 2025, page 87 / 118 26 / 53 flexible and configured to engage and hold tissue in place during processing and embedding. Furthermore, the resilient fingers 42 are capable of successfully sectioning in the microtome after the recess or inner area of ​​the cassette is loaded with liquefied embedding material that subsequently hardens. The resilient fingers 42 can, for example, be formed from the same material as the cap 18, such as a sectionable plastic.

[00070] With reference now to FIGS. 3B and 3C, assembly 10 is shown with the peripheral part 16 in the closed position and where the cassette 12 and the cover 18 are in a first position. Once the fabric 44 is loaded into the interior or recess 11 of the cassette 12, the peripheral part 16 can be rotated to the closed position. The peripheral part 16 rotates around the hinges 22a, 22b, which may be frangible, to move from the open position to the closed position. If the hinges 22a, 22b are frangible, such articulation can move the hinges 22a, 22b away. The peripheral part 16 can rotate until the latches 24, 26 engage with the flanges 30, 32 of the frame 14, securely locking the peripheral part 16 to the frame 14. With the peripheral part 16 in the closed position, the resilient fingers 42 tilt the tissue sample 44 towards the lower wall 12b of the cassette 12.

[00071] As further shown in FIGS. 3B and 3C, when the lid 18 is closed, the resilient fingers 42 press against the tissue sample 44 and deform three-dimensionally around the tissue sample 44, creating three-dimensional spaces around the tissue sample 44 and essentially immobilizing the tissue sample 44 during tissue processing and embedding procedures. This also ensures that the tissue sample 44 is held flat against the lower wall 12b of the cassette 12, so that when slices are made in the microtome, progressively from the lower wall 12b towards the lid 18, complete and continuous sections of tissue sample 44 can be formed. Once the entire sample 44 has been cut, the next slice will contain only the resilient structure 42 and Petition 870250039567, dated 05 / 15 / 2025, pages 88 / 118 27 / 53 Paraffin wax included.

[00072] Now with reference to FIGS. 3D and 4C, assembly 10 is shown in which cassette 12 and cover 18 are in the second position. Pressing cover 18 down causes the cover-frame connectors 36 and the frame-cassette connectors 64 to separate (as described above), allowing cover 18 and cassette 12 to move from the first position (FIG. 3C) to the second position (FIG. 3D). The continued downward pressure on the cover 18 causes the cover 18 to slide further downward into the frame 14. During this downward movement, each corner of the cover 18 (e.g., retaining flanges 38) engages with its respective cassette positioning element 40 on the inner corner of the frame 14. As the retaining flanges 38 pass over the cassette positioning elements 40, they deform the cassette positioning elements 40 and finally “snap” under the cassette positioning elements 40, as shown in FIG.4C, which prevents the upward movement of cap 18 and cassette 12. When cassette 12 reaches the second position, stops 46 prevent the downward movement of flanges 38 of cap 18. This keeps cassette 12 and cap 18 in the second position during embedding and the subsequent sectioning process in the microtome.

[00073] In the second position, the tissue sample 44, a portion of the cassette 12, and resilient finger portions 42 are prepared for sectioning in a microtome. Because the cassette positioning elements 40 and the stop 46 limit the displacement of the lid 18 in the second position, the cassette positioning elements 40 and the stop 46 ensure that the cassette 12 is positioned at a predetermined depth independent of the cassette 12 configuration. The staggering at the predetermined depth ensures that the lower wall 12b of the cassette 12 is positioned as desired in relation to the embedding mold 52, such as at a predetermined vertical spacing (see FIG. 3D). This facilitates the use of microtomes. Petition 870250039567, dated 05 / 15 / 2025, pages 89 / 118 28 / 53 automated in the processes described below, because the thickness of the embedding material, which must be removed before reaching the tissue sample 44 retained against the lower wall 12b, will be substantially the same for all cassettes.

[00074] As millions of procedures are completed each year using such assemblies, embodiments of the present invention are designed for high production volumes and, consequently, are directed to use in automated histopathological processes. One such process is automated embedding. An exemplary automated embedding machine uses a motorized scaling device 48 that pushes the cassette through the frame into the embedding mold 52, as shown in FIGS. 3C and 3D. A scaling device 48 may incorporate spring-loaded cylindrical fingers or feet that push the lid 18 and the cassette 12 through the frame 14.

[00075] In use, one or more tissue samples 44 are placed within the internal space or recess and, specifically, on the lower wall 12b of the cassette 12, as shown in FIG. 3A. The tissue sample 44 is sized and oriented in the cassette 12 according to the required cutting plane desired by the pathologist for each tissue sample 44. The peripheral part 16 is then closed and snapped into place so that the resilient fingers 42 support and clamp the tissue sample 44 against the lower wall 12b in the desired orientation, as shown in FIG. 3B. The resilient fingers 42 can deform three-dimensionally to accommodate various sizes and shapes of tissue samples 44. The force of the resilient fingers 42 against the tissue sample 44 must be sufficient to immobilize the tissue sample 44 and clamp it against the lower wall 12b, but not sufficient to induce artifacts in the tissue sample 44.At this point, assembly 10 with the trapped tissue sample 44 can be subjected to a conventional tissue processing operation that uses vacuum, heat, and chemicals to remove fluids. Petition 870250039567, dated 05 / 15 / 2025, pages 90 / 118 29 / 53 interstitial fluids from within the tissue and replace these fluids with a hardenable material, such as molten paraffin. As mentioned above, during these processing steps, the resilient fingers 42 and the lower wall 12b allow the fluids to reach and fully infiltrate the tissue sample 44.

[00076] The illustrated configuration of the cassette 12, frame 14, and lid 18, including the resilient fingers 42, is an improvement over assemblies requiring a complex lid adjustment procedure, whereby the user must choose from a limited number of specific engagement distances between the lid and the cassette to ensure that the tissue sample 44 is properly immobilized against the lower wall 12b of the cassette. The specific engagement distances were determined by predefined tabs inside the cassette basket that engaged and retained the lid. By using the illustrative embodiment, and particularly the resilient fingers 42 (or other similar tissue propensity structures described herein), the complex adjustment procedure for the lid is eliminated.Consequently, in some exemplary embodiments, lid closure and proper immobilization of the tissue sample (without excessive deformation) can be achieved by a simple “one-hook” procedure in which the lid is closed and the resilient fingers 42 (or other similar propensity structures) properly tilt the tissue sample 44 against the lower wall 12b of the cassette 12, regardless of the thickness of the tissue sample. It will be noted that other configurations and designs can be used to achieve similar purposes.

[00077] After tissue processing is complete, cassette 12 and frame 14 are then placed in a suitable mold 52 and immersed in paraffin. Cassette 12 and / or frame 14 may include machine-readable symbols that allow a machine to determine the type and size of cassette 12 to be used and to make an appropriate decision as to which mold to place in cassette 12 for Petition 870250039567, dated 05 / 15 / 2025, pages 91 / 118 30 / 53 Inclusion. The entire assembly 10, including the exposed portion of cassette 12, is embedded in a hardened block of paraffin wax. The mold 52 may generally follow the contour of the bottom 12b of cassette 12, although the portion of the mold surrounding cassette 12 is preferably square rather than round. This aids in the subsequent production of strip pieces. This part of the procedure may therefore be similar to that described in the patent properties incorporated above. As discussed herein, the frame 14 is then used as a fixing accessory to mount the included assembly 10 in a microtome. The required number of slices are taken from the exposed underside until sufficient sections or slices are taken and properly mounted on a microscope slide, stained, and covered by sliding.

[00078] Another method (not shown) of loading tissue sample 44 into the assembly 10 is possible. First, the peripheral part 16 is detached from the frame 14, which is set aside. The tissue sample 44 is placed on the resilient fingers 42, and then the frame 14 is installed on top of the cover 18. When the frame 14 is installed on top of the peripheral part 16, the latches 24, 26 of the peripheral part 16 engage with the flanges 30, 32, respectively, of the frame 14. In this way, the peripheral part 16 is secured to the frame 14. The assembly 10 can then be positioned in its usual vertical position while the resilient cover 18 remains attached to the frame 14 and the resilient fingers 42 hold the tissue sample 44 to the lower wall 12b of the cassette 12.

[00079] Generally, the cassette 12 (and, in some embodiments, the cover 18) can be formed from a relatively less rigid material, and the frame 14 (and, in some embodiments, the cover 18) can be formed from a relatively more rigid material. The cassette 12 can be formed from a sectionable plastic, such as perfluoroalkoxyethylene (PFA), or materials based on or containing polyethylene (PE), according to the patent properties incorporated above. The material forming the cassette 12 can be at least Petition 870250039567, dated 05 / 15 / 2025, pp. 92-118 31 / 53 translucent, so as not to attract attention during tissue analysis. The frame 14, including the peripheral part 16, can be formed from a more rigid and less expensive plastic, such as acetal. Acetal is much easier to mold in large quantities or in multi-cavity injection molds. As will be observed from FIG. 1A, the cassette 12 can be molded separately from the frame 14 and then inserted into the frame 14. Similarly, the cap 18 can be molded separately from the peripheral part 16 and then inserted into the peripheral part 16. Furthermore, when the cassette 12 and the frame 14 are made of materials with significantly different melting temperatures, they can be molded or co-molded by insertion, such as using the two-injection technique described below in relation to FIGS. 6A-6D.In some illustrative embodiments, cassette 12, cover 18 and frame 14 can be combined into a single piece before the customer receives them, so that the assembly arrives as a single piece ready to load with fabric. This is advantageous compared to previous assemblies, in which the user was required to assemble the components before loading fabric.

[00080] FIG. 5E shows the 36-pin connector, which is structurally similar to the 64-pin connector, but in a generally reversed configuration.

[00081] FIGS. 5A-5D illustrate an example of an assembly process, specifically relating to the cassette frame connectors 64. FIG. 5A shows the frame 14 and the components integrally formed with it, including the tab 70 and the pin 66. Notably, the pin 66 does not yet include its wide end 74 (FIGS. 5C and 5D). The pin 66 can generally be in the form of a truncated cone, which can facilitate the release of the mold used to form the frame 14 and associated components. FIG. 5B shows the cassette and the components integrally formed with it, including the flange 68 positioned in relation to the pin 66, so that the axis 76 of the pin is Petition 870250039567, dated 05 / 15 / 2025, pp. 93 / 118 32 / 53 extend through flange 68. This can be accomplished by mounting a separately molded frame 14 and cassette 12, or it can be accomplished by sequentially molding frame 14 and cassette 12 in place. FIG. 5B also shows tool 98 approaching pin 66. FIG. 5C shows pin 66 with the distal part of its shaft 76 formed into a relatively wider tip 74 by the operation of tool 98. This deformation of pin 66 to form tip 74 fastens flange 68 of cassette 12 to pin 66 of frame 14. In some exemplary processes, tip 74 can be formed by tool 98 when pin 66 is at a temperature above ambient temperature but below its melting temperature, so that the distal part of shaft 76 can be readily plastically deformed by the tool. FIG. Figure 5D shows the complete 64 frame-cassette connectors, as described previously.A corresponding, generally similar process can be used to assemble cover-frame connectors 36.

[00082] FIGS. 6A-6D, 7A-7C and 7D-7F illustrate alternative examples of retaining structures comprising pin / flange connectors and associated assembly methods. The examples described above in relation to FIGS. 5A-5E, or any of the alternative examples described below in relation to FIGS. 6A-6D, 7A-7C and 7D-7F, can be used in combination with various embodiments according to the present description. FIGS. 6A-6D are a cross-sectional view of an alternative pin / flange connector 600 showing the steps of a two-injection co-molding process. In FIG. 6A, a pin 602 is integrally formed with a first component 604, such as by injection molding. This operation can be referred to as a first molding operation or a first injection. In the context of the assembly 10 illustrated in FIG. 1, the first component 604 can be the frame 14. In FIG. 6B, a mold / tool ​​612 is shown approaching the first component 604. With reference to FIG.6C, the mold / tool ​​612. Petition 870250039567, dated 05 / 15 / 2025, pages 94 / 118 33 / 53 engages the first component 604 to deform the pin 602 to form a wider mushroom-head shaped tip 614. In some exemplary processes, the tip 614 can be formed by the mold / tool ​​612 when the pin 602 is at a temperature above ambient temperature but below its melting temperature, so that the distal part of the pin can be readily plastically deformed by the mold / tool. Additionally, the mold / tool ​​612 engages the first component 604 to define at least partially one or more cavities 610 for molding a second material. As shown in FIG. 6D, the cavity 610 is loaded in a second molding operation or second injection to form a flange 606, integrally formed with a second component 608, arranged around the pin 602.Generally, because flange 606 extends into the recess or undercut below the tip and encircles pin 602, the flange is held in place until connector 600 is separated (e.g., broken) during the stepping operation. In the context of assembly 10 illustrated in FIG. 1, the second component 608 could be cassette 12. Such a two-injection molding process can be advantageous compared to some other potential assembly or molding processes because it may involve fewer handling and assembly steps, for example. In an exemplary two-injection process, the first component 604 can be formed in a mold. Without removing the first component 604 from the mold, the mold can be reconfigured (e.g., rotated 90 degrees) to receive the mold / tool ​​612. Then, the second component 608 can be molded. Finally, the first component 604 and the second component 608, now connected, can be removed.Therefore, this exemplary two-injection process may not require the transfer of the first component from a first mold to a second mold or set of separately molded components.

[00083] In other exemplary processes, the first component Petition 870250039567, dated 05 / 15 / 2025, pages 95 / 118 34 / 53 604 can be formed and then placed in a second mold. Then, the second component 608 can be formed, such as by injection molding, directly onto the first component 604, as by a co-molding or insert molding process. Generally, the first component 604 can be shaped so that any cavities generally narrow with increasing depth to facilitate mold release. Similarly, pin 602 can generally be in the shape of a truncated cone, with the narrower end facing outwards, to facilitate mold release.

[00084] FIGS. 7A-7C are a cross-sectional view of an alternative pin / flange connector 700 showing steps in an assembly process. In FIG. 7A, a first component 702 includes a recess 704 in which a pin 706 is installed and secured. For example, the pin 706 can be secured in the recess 704 using an adhesive. The first component 702 can be formed from a first material, and the pin 706 can be formed from a second material. The pin 706 can generally be in the shape of a truncated cone and includes a recess 708 at its distal end. In FIG. 7B, a tool 710 is forming a tip 712 on the pin 706. The tool 710 includes a central projection 714 arranged to engage in the recess 708 of the pin 706. In FIG. 7C, a second component 716 (for example, comprising a flange) was molded into the recess 704 around the pin 706, such as by injection molding. The second component 716 may be formed from a third material.Therefore, connector 700 couples the first component 702 and the second component 716. More specifically, because the flange of the second component 716 extends into the recess or undercut below the tip 712 and encircles the pin 706, the second component is secured to the first component 702 until connector 700 is separated (e.g., broken) during the scaling operation.

[00085] FIGS. 7D-7F are seen in cross-section of a Petition 870250039567, dated 05 / 15 / 2025, pages 96 / 118 35 / 53 Alternative pin / flange connector 750 showing the steps of an assembly process. The pin connector 750 can generally be similar to the pin connector 700, except that the pin 756 can be integrally formed with the first component 752. In FIG. 7D, the first component 752 includes a recess 754 within which a pin 756 is integrally formed. The first component 752 and the pin 756 can be formed from a first material. The pin 756 can generally be in the shape of a truncated cone. In FIG. 7B, a tool 760 is forming a tip 762 on the pin 756. Alternatively, in some exemplary embodiments with appropriate geometries, the tip 762 can be shaped during the initial forming of the first component 752, so that the tool 760 and the separate tip forming step can be omitted. In FIG.7F, a second component 766 (e.g., comprising a flange) was molded into the recess 754 around the pin 756, such as by injection molding in a second mold. The second component 766 may be formed from a second material. Consequently, the connector 750 couples the first component 752 and the second component 766. More specifically, because the flange of the second component 766 extends into the recess or undercut, below the tip 762, and encircles the pin 756, the second component is fixed to the first component 752 until the connector 750 is separated (e.g., broken) in the scaling operation.

[00086] Although several exemplary processes for forming and assembling certain cassette-frame connectors, cover-frame connectors, and other exemplary components are described in this document, it should be understood that various devices, including such connectors and methods of using the devices, may fall within the scope of this description, regardless of whether the devices are produced using the processes described above, conventional processes, or processes developed in the future, or any combination thereof. Petition 870250039567, dated 05 / 15 / 2025, pages 97 / 118 36 / 53

[00087] FIGS. 8, 9 and 10A-10C represent another illustrative embodiment of an assembly 100 that is generally similar to the assembly 10 shown and described in relation to FIGS. 1, 1A, 2, 3A-3D and 4A-4C. Similar reference numbers refer to similar structures shown and described above. Unless specifically indicated, the description of the structure and function or methodology of corresponding components in relation to assembly 10 generally applies to assembly 100. Therefore, repeated explanation of the structure and function or methodology described previously is not necessary. In this embodiment, the resilient fingers 42 of assembly 10 have been replaced by a generally rectangular plate 142 and protrusion members 120. In addition, assembly 100 includes a different arrangement of lid closure elements and cassette closure elements.

[00088] Assembly 100 includes a tissue sample cassette 112 including a recess or internal area 111 surrounded by at least one side wall 112a and including a bottom wall 112b. The cassette 112 is carried within and separably coupled to a frame 114, which includes a peripheral part 116. A lid 118 is separably coupled to the peripheral part 116. The peripheral part 116 generally includes a defined interior between the surrounding (peripheral) walls 116a, 116b, 116c, 116d, and the lid 118 is sized and configured to fit within and is separably coupled to at least one of the surrounding walls 116a, 116b, 116c, 116d. Frame 114 generally includes a defined interior between the surrounding outer walls 114a, 114b, 114c, 114d and a lower edge 114e, and the cassette 112 is sized and configured to move within the interior between at least the first and second positions, as generally described above in relation to mounting 10 and for the same purposes. The first position is shown in FIG. 10B, while the second “stepped” position is shown in FIG. 10C, where the lower part of the cassette 112 is exposed. Petition 870250039567, dated 05 / 15 / 2025, pages 98 / 118 37 / 53 below the lower edge 114e of the frame 114 to allow the cassette 112 and the embedded tissue sample to be sectioned in a microtome while the frame 114 is held in the microtome chuck.

[00089] The connection of the fabric cassette 112 to the frame 114 can be carried out in many different ways, such as any of the ways described above. In the illustrative embodiment of FIG. 8, the cassette 112 is initially separably coupled to the frame 114 by means of frame-cassette connectors 164, which couple the surrounding walls 114a, 114b, 114c, 114d to the cassette 112. The frame-cassette connectors 164 are similar in construction and operation to the frame-cassette connectors 64 described above in relation to assembly 10. In this illustrative embodiment, the frame-cassette connectors 164 are frangible.

[00090] The connection of the cover 118 to the peripheral part 116 of the frame 114 can be carried out in many different ways, such as any of the ways described above. In the illustrative embodiment of FIG. 8, the cover 118 is initially separably coupled to the peripheral part 116 of the frame 114 by means of cover-frame connectors 136, which couple the surrounding walls 116a, 116b, 116c, 116d to the cover 118. The cover-frame connectors 136 are similar in construction and operation to the cover-frame connectors 36 described above in relation to assembly 10. In this illustrative embodiment, the cover-frame connectors 136 are frangible.

[00091] Now with reference to FIGS. 8 and 9, the connections between the frame 114 and the peripheral part 116 are shown in more detail. The peripheral part 116 is coupled to the wall 114a of the frame 114 by a pair of hinges 122a, 122b, which are optionally frangible. The peripheral part 116 snaps into the closed position (FIG. 10B) by engaging closing elements of the peripheral part, such as latches 124, 126, with frame closing elements, such as flanges 130, 132. The latch 124 is positioned on the outer wall. Petition 870250039567, dated 05 / 15 / 2025, pages 99 / 118 38 / 53 116a of the peripheral part 116 and engages with flange 130 on wall 114a of frame 114 in the closed position. The latches 126 are positioned on wall 116c of the peripheral part 116 and engage with flange 132 of wall 114c of frame 114 in the closed position.

[00092] With reference to FIGS. 8, 9 and 10A, the connections between the cover 118 and the cassette 112 are shown in more detail. The cover 118 is press-fitted into the closed position (FIG. 10B) by means of the engagement of cover locking elements, which may include connectors such as latches 158, with cassette locking elements, which may include connectors such as flanges 160. In this embodiment, each latch 158 is disposed on a respective extension arm 159. The arms 159 generally extend downwards from the cover 118 when the cover 118 is in the closed position. A corresponding pair of arms 159 and latches 158 are arranged on each lateral side of the cover 118, with the arms 159 and latches 158 in each pair arranged on the cover in an opposite arrangement and spaced apart, so that the latches 158 face each other.Each flange 160 of the cassette 112 is generally oriented laterally and is arranged to engage with both latches 158 of a corresponding pair of arms 159. In the closed position, with the latches 158 of the lid 118 engaged with flanges 160 of the cassette 112, the lid 118 and the cassette 112 are coupled together and move between the first position (FIG. 10B) and the second position or staggered position (FIG. 10C) as a single unit. Generally, the lid closing elements serve to secure the lid to the cassette and to prevent the lid and cassette from separating after the lid has been closed onto the cassette, such as during various processing operations. In some exemplary embodiments, including lid-mounted components arranged to tilt the tissue sample against the lower wall of the cassette, the lid closing elements may be configured to withstand the corresponding reaction force that may tend to separate the lid from the cassette. Petition 870250039567, dated 05 / 15 / 2025, pp. 100 / 118 39 / 53

[00093] The cassette 112 and the cover 118 are dimensioned and configured to move within the interior of the frame 114 between at least the first and second positions, as best shown in FIGS. 10B and 10C, in a manner similar to that described above in relation to the assembly 10. The cover 118 includes a cover retaining flange 138 at each of its four corners. The retaining flanges 138 are configured to engage with cassette positioning elements 140 of the frame 114, which are formed as part of the inner corners of the four corners of the frame 114. In the illustrative embodiment, each retaining flange 138 engages with a respective cassette positioning element 140.The cassette positioning elements 140 are flexible and hollow, so that as the retaining flanges 138 pass through the cassette positioning elements 140 (e.g., downwards), the retaining flanges 138 deform the cassette positioning elements 140 and finally “snap” under the cassette positioning elements 140, as shown, for example, in FIG. 10C.

[00094] As shown in FIGS. 9 and 10B, each corner of the frame 114 includes a diagonal stop 146 located below and away from the descending part of the respective cassette positioning element 140. When the cassette 112 reaches the second position, the stops 146 prevent the downward movement of the retaining flanges 138 of the lid 118. Thus, in the second position, the retaining flanges 138 of the lid 118 are fixed vertically between the cassette positioning elements 140 (at the top) and the stops 146 (at the bottom). As the lid 118 and the cassette 112 are coupled together by latches 158 and flanges 160, this keeps the cassette 112 and the lid 118 in the second position and ready for embedding and subsequent sectioning in the microtome, as described above.

[00095] With reference to FIGS. 8, 9 and 10A-10C, instead of the resilient fingers 42 of assembly 10 described above, the cover 118 of this embodiment includes a resilient structure comprising a plate Petition 870250039567, dated 05 / 15 / 2025, pages 101 / 118 40 / 53 142 configured to be received within the generally rectangular recess 111 of the cassette 112. The plate 142 is generally rectangular and is coupled to a peripheral part 117 of the cover 118 by a plurality of protrusion members 120. The protrusion members 120 are arranged to deflect the plate 142 towards the lower wall 112b of the cassette 112 when the cover 118 is in the closed configuration. In this embodiment, the protrusion members 120 are arranged in a generally perpendicular arrangement between the plate 142 and the peripheral part 117 of the cover 118.

[00096] Generally, the protrusion members 120 are elastically deformable to allow flexible engagement between the plate 142 and one or more tissue samples in the recess 111 of the cassette 112. The plate 142 and the protrusion members 120 form a compliant structure that holds the tissue in the desired orientation without creating an artifact impression on the tissue sample during processing. Although a generally rectangular recess 111 is shown, it will be noted that any other shape, such as any polygon (e.g., square) or any rounded shape (e.g., oval or circular) or shapes with depression or alignment features for the tissue sample, may be used in its place.

[00097] Plate 142 allows the infiltration of solvents and chemicals used to fix, process, and stain tissue, and of the embedding material used to incorporate the tissue while the tissue is retained by Plate 142. Plate 142 is flexible and configured to engage and hold the tissue in place during processing and embedding. Furthermore, Plate 142 is capable of successfully sectioning in the microtome after the recess 111 or the inner area of ​​the cassette is loaded with liquefied embedding material that subsequently hardens. Plate 142 can, for example, be formed from the same material as the lid 118, such as a sectionable plastic.

[00098] With reference to FIG. 10A, in this embodiment, the side of Petition 870250039567, dated 05 / 15 / 2025, pages 102 / 118 41 / 53 contact with the tissue plate 142 includes teeth 142b, which can generally extend downwards when the cover 118 is in the closed position. The teeth 142b can provide additional security against unwanted movement of the tissue samples.

[00099] Generally, the assembly 100 is loaded with tissue, moved from the open to the closed position (including breaking frangible cap-frame connectors 136 and cassette frame connectors 164), stepped, and otherwise used in a manner similar to the assembly 10 described above. In use, one or more tissue samples are placed within the internal space or recess 111 and, specifically, on the lower wall 112b of the cassette 112. The tissue sample is sized and oriented in the cassette 112 according to the cutting plane desired by the pathologist. The peripheral part 116 is then closed and snapped into place such that the plate 142 supports and holds the tissue sample against the lower wall 112b in the desired orientation. The protrusion members 120 deform to allow the plate 142 to accommodate various sizes and shapes of tissue samples.The force of plate 142 against the tissue sample must be sufficient to immobilize the tissue sample, but not sufficient to induce artifacts in the tissue sample. At this point, mount 100, with the tissue sample clamped, can be subjected to a conventional tissue processing operation that uses vacuum, heat, and chemicals to remove interstitial fluids from within the tissue and replace these fluids with a hardenable material, such as molten paraffin. As mentioned above, during these processing steps, plate 142 and the bottom wall 112b allow fluids to fully reach and infiltrate the tissue sample. Furthermore, plate 142 clamps the tissue sample flat against the bottom wall 112b without leaving artifacts or markings on the tissue that could interfere with subsequent microscopic analysis.It will be observed that different materials and configurations of the protrusion member 120 and the plate 142 can be chosen based, for example, on the type of. Petition 870250039567, dated 05 / 15 / 2025, pages 103 / 118 42 / 53 tissue to be processed and analyzed. For example, small samples of mucosal tissue can be successfully maintained and processed using some arrangements, while other tissue types, such as adipose tissue, may be better served by another material or configuration. For example, the thickness, shape, and number of protrusion limbs can be selected to provide a desired immobilization force on the tissue samples. [000100] After the tissue processing is complete, cassette 112 and frame 114 are then placed in a suitable mold and immersed in paraffin. Cassette 112 and / or frame 114 may include machine-readable symbols, allowing a machine to determine the type and size of cassette 112 to be used and to make an appropriate decision as to which mold to place cassette 112 in for embedding, generally in the manner described above with reference to assembly 10. [000101] FIGS. 11, 12, 13A, 13B and 14A-C represent another illustrative embodiment of an assembly 200 that is similar to assembly 100 shown and described in relation to FIGS. 8, 9 and 10A-10C, as well as assembly 10. The similar reference numbers in FIGS. 11, 12, 13A, 13B and 14A-C refer to the similar structures shown and described above. Unless specifically indicated, the description of the structure and function or methodology of corresponding components in relation to assembly 100 and assembly 10 generally applies to assembly 200. Therefore, repeated explanation of the structure and function or methodology described above is not necessary. In this embodiment, the generally rectangular plate 142 of assembly 100 was replaced by the generally circular plate 242, and the generally perpendicular pretension members 120 of assembly 100 were replaced by generally helical pretension members 220.In addition, the 200 assembly includes a different arrangement of lid closing elements and cassette closing elements. [000102] Assembly 200 includes a tissue sample cassette Petition 870250039567, dated 05 / 15 / 2025, pages 104 / 118 43 / 53 212 including a recess or internal area 211 surrounded by at least one side wall 212a and including a bottom wall 212b. The cassette 212 is carried within and separably coupled to a frame 214, which includes a peripheral part 216. A cover 218 is separably coupled to the peripheral part 216. The peripheral part 216 generally includes a defined interior between the surrounding (peripheral) walls 216a, 216b, 216c, 216d, and the cover 218 is dimensioned and configured to fit the interior and is separably coupled to at least one of the surrounding walls 216a, 216b, 216c, 216d. Frame 214 generally includes a defined interior between the surrounding outer walls 214a, 214b, 214c, 214d and a lower edge 214e, and the cassette 212 is sized and configured to move within the interior between at least the first and second positions, as generally described above in relation to mounting 100 and for the same purposes. The first position is shown in FIG. 14B, while the second “stepped” position is shown in FIG. 14C, wherein the lower part of the cassette 212 is exposed below the lower edge 214e of the frame 214 to allow the cassette 212 and the embedded tissue sample to be sectioned in a microtome while the frame 214 is held in the microtome chuck. [000103] The connection of the fabric cassette 212 to the frame 214 can be accomplished in many different ways, such as any of the ways described above. In the illustrative embodiment of FIG. 11, the cassette 212 is initially separably coupled to the frame 214 via frame-cassette connectors 264 that couple the surrounding walls 214a, 214b, 214c, 214d to the cassette 212. The frame-cassette connectors 264 are similar in construction and operation to the frame-cassette connectors 164 described above in relation to assembly 100. In this illustrative embodiment, the frame-cassette connectors 264 are frangible. [000104] The connection of cover 218 to the peripheral part 216 of the frame Petition 870250039567, dated 05 / 15 / 2025, pages 105 / 118 44 / 53 214 can be implemented in many different ways, such as any of the ways described above. In the illustrative embodiment of FIG. 11, the cover 218 is initially separably coupled to the peripheral part 216 of the frame 214 by means of cover-frame connectors 236 which couple the surrounding walls 216a, 216b, 216c, 216d to the cover 218. The cover-frame connectors 236 are similar in construction and operation to the cover-frame connectors 136 described above in relation to assembly 100. In this illustrative embodiment, the cover-frame connectors 236 are frangible. [000105] Now with reference to FIGS. 11, 12 and 13A, the connections between the frame 214 and the peripheral part 216 are shown in more detail. The peripheral part 216 is coupled to the wall 214a of the frame 214 by a pair of hinges 222a, 222b, which are optionally frangible. The peripheral part 216 snaps into place in the closed position (FIG. 14B) by engaging the peripheral part's locking elements, such as latches 224, 226, with the frame's locking elements, such as flanges 230, 232. The latch 224 is positioned on the outer wall 216a of the peripheral part 216 and engages with flange 230 on the wall 214a of the frame 214 in the closed position. The latches 226 are positioned on wall 216c of the peripheral part 216 and engage with a flange 232 of wall 214c of frame 214 in the closed position. [000106] With reference to FIGS. 11, 12 and 14A, the connections between the cover 218 and the cassette 212 are shown in more detail. The cover 218 is press-fitted into the closed position (FIG. 14B) by means of the engagement of cover locking elements, which may include connectors such as latches 258, with cassette locking elements, which may include connectors such as flanges 260. In this embodiment, each of the four latches 258 is arranged on a respective extension arm 259. The arms 259 generally extend downwards from the cover 218 when the cover 218 is in the closed position. The arms 259 are arranged in an arrangement Petition 870250039567, dated 05 / 15 / 2025, pages 106 / 118 45 / 53 spaced apart generally encircling plate 242, and the latches 258 are generally oriented laterally outwards from their respective arms. Each of the four flanges 260 of cassette 212 is generally oriented laterally and is arranged to engage a latch 258. In the closed position, with the latches 258 of cover 218 engaged with the flanges 260 of cassette 212, cover 218 and cassette 212 are coupled and move between the first position (FIG. 14B) and the second or staggered position (FIG. 14C) as a single unit. [000107] The cassette 212 and the cover 218 are dimensioned and configured to move within the interior of the frame 214 between at least the first and second positions, as shown best in FIGS. 14B and 14C in a manner similar to that described above in relation to the assembly 100. The cover 218 includes a cover retaining flange 238 at each of its four corners. The retaining flanges 238 are configured to engage with the cassette positioning elements 240 of the frame 214, which are formed as part of the inner corners of the four corners of the frame 214. In the illustrative embodiment, each retaining flange 238 engages with a respective cassette positioning element 240.The cassette positioning elements 240 are flexible and hollow, so that as the retaining flanges 238 pass through the cassette positioning elements 240 (e.g., downwards), the retaining flanges 238 deform the cassette positioning elements 240 and finally “snap” under the cassette positioning elements 240, as shown, for example, in FIG. 14C. [000108] As best shown in FIG. 14A, each corner of the frame 214 includes a diagonal stop 246 located below and away from the downward-sloping part of the respective cassette positioning element 240. When the cassette 212 reaches the second position, the stops 246 prevent the downward movement of the retaining flanges 238 of the cover 218. Thus, in the second position, the retaining flanges 238 of the cover 218 are vertically fixed between Petition 870250039567, dated 05 / 15 / 2025, pages 107 / 118 46 / 53 the cassette positioning elements 240 (at the top) and the stops 246 (at the bottom). As the lid 218 and the cassette 212 are coupled together by latches 258 and flanges 260, this keeps the cassette 212 and the lid 218 in the second position and ready for embedding and subsequent sectioning in the microtome, as described in the patent properties incorporated above. [000109] With reference to FIGS. 11, 12, 13A, 13B and 14A-14C, instead of the generally rectangular plate 142 of the assembly 100 described above, the cover 218 of this embodiment includes a resilient structure comprising a generally circular plate 242 configured to be received within the generally circular recess 211 of the cassette 212. The plate 142 is coupled to a peripheral part 217 of the cover 218 by a plurality of protrusion elements 220. The protrusion members 220 are arranged to tilt the plate 242 towards the lower wall 212b of the cassette 212 when the cover 218 is in the closed configuration. In this embodiment, the protrusion members 220 are arranged in an angled arrangement and, more specifically, a generally helical arrangement between the plate 242 and the peripheral part 217 of the cover 218. [000110] Generally, the propensity members 220 are elastically deformable to allow flexible engagement between the plate 242 and one or more tissue samples in the recess 211 of the cassette 212. The plate 242 and the propensity members 220 form a compliant structure that holds the tissue in the desired orientation without creating an artifact impression on the tissue sample during processing. Although a generally circular recess 211 is shown, it will be noted that any other shape, such as any polygon (e.g., square or rectangle) or any rounded shape (e.g., oval) or shapes with depression or alignment features for the tissue sample, may be used in its place. [000111] Plate 242 allows infiltration of solvents and products Petition 870250039567, dated 05 / 15 / 2025, pages 108 / 118 47 / 53 chemicals used to fix, process, and stain the tissue, and the embedding material used to incorporate the tissue while the tissue is retained by plate 242. Plate 242 is flexible and configured to engage and hold the tissue in place during processing and embedding. Furthermore, plate 242 is capable of successfully sectioning in the microtome after recess 211 or the inner area of ​​the cassette is loaded with liquefied embedding material that subsequently hardens. Plate 242 can, for example, be formed from the same material as the lid 218, such as a sectionable plastic. [000112] With reference to FIG. 14A, in this embodiment, the tissue contact side of plate 242 includes teeth 242b, which can generally extend downwards when the lid 218 is in the closed position. The teeth 242b can provide additional security against unwanted movement of the tissue sample. [000113] Generally, the 200 mount is loaded with tissue, moved from the open to the closed position (including breaking frangible cap-frame connectors 236 and cassette frame connectors 264), staggered, and otherwise used in a manner similar to the 100 and 10 mounts described above. In use, one or more tissue samples are placed within the internal space or recess 211 and, specifically, over the lower wall 212b of the cassette 212. The tissue sample is sized and oriented in the cassette 212 according to the cutting plane desired by the pathologist. The peripheral part 216 is then closed and snapped into place such that the plate 242 supports and holds the tissue sample against the lower wall 212b in the desired orientation. The protrusion members 220 may deform to allow the plate 242 to accommodate various sizes and shapes of tissue samples.The force of plate 242 against the tissue sample must be sufficient to immobilize the tissue sample, but not sufficient to induce artifacts in the tissue sample. At this point, mount 200 with the clamped tissue sample can be subjected to a conventional tissue processing operation using vacuum. Petition 870250039567, dated 05 / 15 / 2025, pages 109 / 118 48 / 53 Heat and chemicals are used to remove interstitial fluids within the tissue and replace these fluids with a hardenable material, such as molten paraffin. As mentioned above, during these processing steps, plate 242 and the bottom wall 212b allow fluids to fully reach and infiltrate the tissue sample. Furthermore, plate 242 holds the tissue sample flat against the bottom wall 212b without leaving artifacts or markings on the tissue that could interfere with subsequent microscopic analysis. It will be observed that different materials and configurations of the propensity member 220 and plate 242 can be chosen based on, for example, the type of tissue to be processed and analyzed. For example, small mucosal tissue samples can be successfully held and processed using some arrangements, while other tissue types, such as adipose tissue, may be better served by another material or configuration.For example, the thickness, shape, and number of 220 propensity members can be selected to provide a desired immobilization force on tissue samples. [000114] After the tissue processing is complete, cassette 212 and frame 214 are then placed in a suitable mold and immersed in paraffin. Cassette 212 and / or frame 214 may include machine-readable symbols, allowing a machine to determine the type and size of cassette 212 to be used and to make an appropriate decision as to which mold to place cassette 212 in for embedding, generally in the manner described above with reference to mounting 100 and mounting 10. [000115] FIGS. 15, 16A, and 16B represent another illustrative embodiment of an assembly 300, which is generally similar to the assembly 10 shown and described in relation to FIGS. 1, 1A, 2, 3A-3D, and 4A-4C, as well as other illustrative embodiments described herein. Similar reference numbers refer to the similar structures shown and described above. Unless specifically indicated, the description of the structure and function or methodology of components Petition 870250039567, dated 05 / 15 / 2025, pp. 110 / 118 49 / 53 corresponding to assembly 10 and other illustrative embodiments generally apply to assembly 300. Therefore, repeated explanation of the structure and function or methodology described previously is not necessary. [000116] In assembly 300, the frame-cover connectors 36 and the cassette-cover connectors 64 of assembly 10 have been replaced by frame-cover connectors 336 and cassette-cover connectors 364, respectively, which generally comprise inverted versions of frame-cover connectors 36 and cassette-cover connectors 64. Furthermore, assembly 300 includes tension lifters 385 on flanges 384. Additionally, assembly 300 includes peripheral part closure elements (e.g., latches 25 and flanges 31) arranged to couple surrounding side (peripheral) walls 316b, 316d to the side surrounding the outer walls 314b, 314d, respectively. Any one or more of these features may optionally be included in any other exemplary embodiment. [000117] Assembly 300 includes a tissue sample cassette 312 generally similar to sample cassette 12. Cassette 312 is carried within and separably coupled to a frame 314, which includes a peripheral part 316 and is generally similar to frame 14 and peripheral part 16. A lid 318 (generally similar to lid 18) is separably coupled to the peripheral part 316. The peripheral part 316 generally includes an interior defined between the surrounding (peripheral) walls 316a, 316b, 316c, 316d, and the lid 318 is sized and configured to fit within the interior and is separably coupled to at least one of the surrounding walls 316a, 316b, 316c, 316d. Frame 314 typically includes a defined interior between the surrounding exterior walls 314a, 314b, 314c, 314d, and the 312 cassette is sized and configured to move inside between at least the first and second positions, as generally described above in relation to mounting 10 and for the same purposes. Petition 870250039567, dated 05 / 15 / 2025, pages 111 / 118 50 / 53 [000118] The connection of the fabric cassette 312 to the frame 314 can be accomplished in many different ways, such as any of the ways described above. In the illustrative embodiment of FIG. 15, the cassette 312 is initially separably coupled to the frame 314 via frame-cassette connectors 364 that couple the surrounding walls 314a, 314b, 314c, 314d to the cassette 312. The frame-cassette connectors 364 are similar in construction and operation to the frame-cassette connectors 64 described above in relation to assembly 10, except that their orientation is reversed. [000119] Specifically, each frame-cassette connector 364 includes a retaining structure (e.g., first retaining structure), such as a pin 366, integrally formed with the frame 314 and extending at least partially through a retaining structure (e.g., second retaining structure), such as a flange 368 integrally formed with the cassette 312. Each pin 366 generally extends downward (compared to generally upward in assembly 10) from a tab 370, which generally extends laterally into one of the outer walls 314a, 314b, 314c, 314d. Generally, because the tip of the pin 366 and the tab 370 are wider than the shaft extending between them and because the flange 368 extends into the recess or recessed area between the tip and the tab, the flange is retained in the pin by the tip and the tab.Consequently, until the cassette-frame connector 364 is separated (e.g., broken) during the scaling operation, the flange 368 is fixed to the pin 366. [000120] The cassette-frame connectors 364 of this illustrative embodiment are frangible and are configured to break when the cassette 312 is moved from the first position to the second position, as described above in relation to mounting 10. In this illustrative embodiment, the pin 366 pulled from the flange 368 breaks the flange 368 during this movement. Each tab 370 can be articulated to its respective outer wall 314a, 314b, 314c, 314d, of Petition 870250039567, dated 05 / 15 / 2025, pages 112 / 118 51 / 53 so that, during the movement from the first position to the second position, the tab 370 articulates downwards, which can promote the predictable and consistent separation (e.g., breakage) of the frame-cassette connector 364 (e.g., pins 366 pulled out of the flanges 368). [000121] The connection of the cover 318 to the peripheral part 316 of the frame 314 can be carried out in many different ways, such as any of the ways described above. In the illustrative embodiment of FIG. 15, the cover 318 is initially separably coupled to the peripheral part 316 of the frame 314 by means of cover-frame connectors 336 which couple the surrounding walls 316a, 316b, 316c, 316d to the cover 318. The cover-frame connectors 336 are similar in construction and operation to the cover-frame connectors 36 described above in relation to assembly 10, except that their orientation is reversed. [000122] Specifically, the cap-frame connectors 336 can generally be similar in structure and operation to the frame-cassette connectors 364 described above. Each cap-frame connector 336 includes a retaining structure (e.g., first retaining structure), such as a pin 382, ​​integrally formed with the peripheral part 316 of the frame 314 and extending at least partially through a retaining structure (e.g., second retaining structure), such as a flange 384 integrally formed with the cap 318. In the open position, each pin 382 generally extends downwards (compared to generally upwards in the assembly 10) from a tab 386, which generally extends laterally into one of the surrounding walls 316a, 316b, 316c, 316d. Similarly, in the closed configuration (e.g., similar to FIG. 3B), pins 382 generally extend upwards from tabs 386.Pin 382 includes a base disposed on flange 386, a tip on an opposite surface of flange 384, and a shaft extending from the base to the tip through flange 384. Petition 870250039567, dated 05 / 15 / 2025, pages 113 / 118 52 / 53 [000123] The cover-frame connectors 336 of this illustrative embodiment are frangible and are configured to break when the cover 318 is moved from the first position to the second position, as described above in relation to assembly 10. In this illustrative embodiment, the pin 382 pulled from the flange 384 breaks the flange 384 during this movement. Each tab 386 can be articulated to its respective surrounding wall 316a, 316b, 316c, 316d, so that, during the movement from the first position to the second position, the tab 386 articulates downwards, which can promote the predictable and consistent separation (e.g., breakage) of the cover-frame connector 336 (e.g., pins 382 pulled from flanges 384). [000124] Any exemplary frangible connectors described herein may include one or more strain boosters arranged to encourage a specific failure mode, such as which component of the connector will break first and / or the location of the break. FIG. 16 illustrates an exemplary strain booster, specifically a notch 385 in the flange 384. During scaling (e.g., movement from the first position to the second position), the notch 385 may promote the breakage of the flange 384, thus facilitating the predictable and consistent separation of the cover 318 from the peripheral part 316 of the frame 314. [000125] Returning to FIG. 15, the peripheral part 316 is press-fitted into the closed position (e.g., similar to FIG. 3B) by engaging the closing elements of the peripheral part, such as latches 25, with frame closing elements, such as flanges 31. In assembly 300, latches 25 extend from the side surrounding the walls 316b, 316d, and the corresponding flanges 31 are arranged on the side surrounding the outer walls 314b, 314d. In various embodiments, the closing elements on the side walls (e.g., latches 25 and flanges 31) may be provided in addition to or in place of the closing element on the end walls (e.g., Petition 870250039567, dated 05 / 15 / 2025, pages 114 / 118 53 / 53 example, latches 24, 26 and flanges 30, 32 of assembly 10). [000126] Although the present invention has been illustrated by the description of its specific embodiments, and although the embodiments have been described in considerable detail, it is not intended to restrict or in any way limit the scope of the claims appended to such details. The various features discussed in this document can be used alone or in any combination within and between the various embodiments. Additional advantages and modifications will readily appear to those skilled in the art. The invention in its broadest aspects is therefore not limited to the specific details, representative apparatus and methods, and illustrative examples shown and described. Consequently, deviations from such details may be made without departing from the scope or the spirit of the overall inventive concept. Petition 870250039567, dated 05 / 15 / 2025, pages 115 / 118

Claims

1 / 6 CLAIMS 1. Histological Tissue Sample Support Device, (10, 100, 200, 300), comprising: a tissue cassette (12, 112, 212, 312) having a recess (11, 111, 211) including at least one side wall (12a, 112a, 212a) and a bottom wall (12b, 112b, 212b), the tissue cassette formed of a first material that can be successfully sectioned in a microtome and is resistant to degradation by solvents and chemicals used to fix, process and stain the tissue; a frame (14, 114, 214, 314) including a lower edge (14e, 114e, 214e), the frame formed of a second material different from the first material and more rigid than the first material, the fabric cassette being coupled to the frame by a frame-cassette connector (64, 164, 264, 364) comprising a first retaining structure formed integrally with the frame extending, at least partially, through a second retaining structure formed integrally with the cassette;and a lid (18, 118, 218, 318) attached to the frame; characterized in that the lid and the tissue cassette are capable of moving from a first position to a second position relative to the frame and, in the second position, the lower wall and at least part of the side wall extend downwards beyond the lower edge of the frame for sectioning in the microtome; and in that the frame is capable of being detached from the cassette by separating the frame-cassette connector.

2. Histological Tissue Sample Support Device, (10, 100, 200, 300), according to Claim 1, characterized by Petition 870230086173, dated 09 / 27 / 2023, page 5 / 11 2 / 6, wherein the first retention structure comprises a pin (66, 366) integrally formed with the frame (14, 114, 214, 314), and the second retention structure comprises a flange (68, 368) integrally formed with the cassette (12, 112, 212, 312).

3. Histological Tissue Sample Support Device, (10, 100, 200, 300), according to Claim 2, characterized in that the flange (68, 368) integrally formed with the cassette (12, 112, 212, 312) is configured to break when the lid (18, 118, 218, 318) and the tissue cassette are moved from the first position to the second position.

4. Histological Tissue Sample Support Device, (10, 100, 200, 300), according to Claim 3, characterized in that the flange (68, 368) integrally formed with the cassette (12, 112, 212, 312) comprises a tension elevator (385) arranged to promote the breakage of the flange integrally formed with the cassette, when the lid (18, 118, 218, 318) and the tissue cassette are moved from the first position to the second position.

5. Histological Tissue Sample Support Device, (10, 100, 200, 300), according to Claim 2, characterized in that the frame (14, 114, 214, 314) further comprises outer walls (14a, 14b, 14c, 14d, 114a, 114b, 114c, 114d, 214a, 214b, 214c, 214d) extending upwards from the lower edge (14e, 114e, 214e); and in that the pin (66, 366) formed integrally with the frame is disposed in a tab (70, 370) that extends laterally into one of the outer walls.

6. Histological Tissue Sample Support Device, (10, 100, 200, 300), according to Claim 5, characterized by Petition 870230086173, dated 09 / 27 / 2023, p. 6 / 11 3 / 6 in that the pin (66, 366) formed integrally with the frame (14, 114, 214, 314) extends downward from the tab (70, 370).

7. Histological Tissue Sample Support Device, (10, 100, 200, 300), according to Claim 5, characterized in that the tab (70, 370) extending from one of the outer walls (14a, 14b, 14c, 14d, 114a, 114b, 114c, 114d, 214a, 214b, 214c, 214d) is articulatedly coupled to one of the outer walls.

8. Histological Tissue Sample Support Device, (10, 100, 200, 300), according to Claim 1, characterized in that the lid (18, 118, 218, 318) is coupled to the frame (14, 114, 214, 314) by a lid-frame connector (36, 136, 236, 336) comprising a third retaining structure formed integrally with a peripheral part (16, 116, 216, 316) of the frame extending, at least partially, through a fourth retaining structure formed integrally with the lid; and wherein the lid is capable of being uncoupled from the peripheral part of the frame, separating the lid-frame connector.

9. Histological Tissue Sample Support Device, (10, 100, 200, 300), according to Claim 8, characterized in that the third retention structure comprises a pin (82, 382) integrally formed with the peripheral part (16, 116, 216, 316) of the frame (14, 114, 214, 314) and the fourth retention structure comprises a flange (84, 384) integrally formed with the cap (18, 118, 218, 318).

10. Histological Tissue Sample Support Device, (10, 100, 200, 300), according to Claim 9, characterized in that moving the lid (18, 118, 218, 318) and the tissue cassette (12, 112, 212, 312) from the first position to the second position breaks the flange formed integrally with the lid. Petition 870230086173, dated 09 / 27 / 2023, page 7 / 11 4 / 6 11. Histological Tissue Sample Support Device, (10, 100, 200, 300), according to Claim 9, characterized in that the peripheral part (16, 116, 216, 316) of the frame (14, 114, 214, 314) comprises peripheral walls (16a, 16b, 16c, 16d, 116a, 116b, 116c, 116d, 216a, 216b, 216c, 216d, 316a, 316b, 316c, 316d) surrounding the lid (18, 118, 218, 318); and wherein the pin (82, 382) formed integrally with the peripheral part is disposed in a tab (86, 386) that extends laterally into one of the peripheral walls.

12. Histological Tissue Sample Support Device, (10, 100, 200, 300), according to Claim 11, characterized in that, when the lid (18, 118, 218, 318) is in a closed configuration, the pin (82, 382) formed integrally with the peripheral part (16, 116, 216, 316) extends below the tab (86, 386).

13. Histological Tissue Sample Support Device, (10, 100, 200, 300), according to Claim 11, characterized in that the tab (86, 386) extending from one of the peripheral walls (16a, 16b, 16c, 16d, 116a, 116b, 116c, 116d, 216a, 216b, 216c, 216d, 316a, 316b, 316c, 316d) is articulatedly coupled to one of the peripheral walls.

14. Method for Preparing One or More Biopsy Tissue Samples for Histological Examination, characterized in that the method comprises: positioning a tissue sample in a tissue cassette (12, 112, 212, 312) having a recess (11, 111, 211) including at least one lateral wall (12a, 112a, 212a) and one inferior wall (12b, 112b, 212b), the tissue cassette formed from a first material that Petition 870230086173, dated 09 / 27 / 2023, page. 8 / 11 5 / 6 can be successfully sectioned in a microtome and is resistant to degradation by solvents and chemicals used to fix, process and stain the tissue, the tissue cassette being disposed in a frame (14, 114, 214, 314) which includes a lower edge (14e, 114e, 214e), the tissue cassette being coupled to the frame by a frame-cassette connector (64, 164, 264,364) which includes a first retention structure formed integrally with the frame extending through a second retention structure formed integrally with the cassette; closing a peripheral part of the frame, the peripheral part of the frame including a cassette cover (18, 118, 218, 318) disposed thereon; moving the cover and the tissue cassette from a first position to a second position relative to the frame, including breaking the frame-cassette connector, wherein, in the second position, the lower wall and at least a part of the side wall extend downwards beyond the lower edge of the frame for sectioning in the microtome.

15. Method for Preparing One or More Biopsy Tissue Samples, according to Claim 14, characterized in that the first retention structure comprises a pin (66, 366) integrally formed with the frame (14, 114, 214, 314) and the second retention structure comprises a flange (68, 368) integrally formed with the cassette (12, 112, 212, 312).

16. Method for Preparing One or More Biopsy Tissue Samples, according to Claim 14, characterized in that the closing operation includes fixing the tissue sample in the recess (11, 111, 211) using a plate (142, 242) mounted on the lid (18, 118, 218, 318) by protrusion elements (120, 220) arranged to tilt the plate towards the lower wall of the recess. Petition 870230086173, dated 09 / 27 / 2023, page 9 / 11 6 / 6

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