JP4584259B2 - Cassettes and procedures for managing and holding tissue specimens during processing, embedding and microtome procedures - Google Patents

Description

[Related applications]
This application claims priority based on U.S. Patent Application Publication No. 60 / 512,147, filed Oct. 17, 2003, and is generally pending, by reference. It relates to the international patent application PCT / US02 / 30779 and the international patent application PCT / US02 / 30775, the entirety of which is incorporated herein.

  The present invention generally relates to a support for handling and embedding a tissue specimen for pathological examination. More particularly, the present invention relates to a cassette that can receive and embed one or more tissue specimens, after which the one or more tissue specimens can be microtomed.

  In order to accurately diagnose various tissue diseases and conditions, medical personnel must remove one or more tissue specimens from the patient's body. This process of removing tissue from the body is known as a biopsy. Once one or more tissue specimens have been removed and sent to a pathology laboratory, the tissue is diagnosed through a series of procedures performed by a histotechnician and ultimately a pathologist. The present invention generally relates to a procedure typically performed by a histologist to prepare one or more tissue cells as a slide that can be analyzed microscopically by the pathologist.

  Although the singular term “specimen” is used throughout this specification, it should be understood that the term includes a plurality of “specimens” as well. When a tissue specimen is removed from a patient's body, it is typically placed in a specimen container that contains a tissue fixative solution, which is then transferred to a pathology laboratory. The tissue undergoes a process known as “grossing-in” in the pathology laboratory. During the process, a histologist removes the tissue specimen from the container, typically cuts the tissue to a size suitable for tissue processing, and places each specimen in a suitably sized small cassette. Place and assign a tracking number to each cassette. The tracking number is then recorded in a tracking system utilized in the laboratory. For minimal tissue specimens, it may be just scraping, but the cassette has precise mesh openings on the sides and bottom wall. In another embodiment comprising a very small tissue specimen, the specimen is placed in a bag similar to a tea bag to prevent the smallest tissue specimen from falling off. Larger tissue specimens are mounted in cassettes that have somewhat larger slot openings than the tissue specimens in the cassette.

  The cassette is then placed inside a stainless steel perforated basket and applied to a tissue processing device. This can often take overnight. The device uses a combination of vacuum, heat, and chemicals to remove tissue fluid. When the fluid is removed from the tissue specimen, the processor immerses in molten paraffin water to replace the tissue fluid in the tissue with paraffin. The histologist then removes the basket from the device and removes the cassette for the individual tissue. The histologist removes tissue from each cassette at an embedding station that individually has a reservoir and a dispenser of molten paraffin. The histologist carefully determines the orientation of the tissue specimen based on the tissue shape within a stainless steel base mold that is approximately the same size as the tissue cassette and partially filled with molten paraffin. Must. The tissue specimen must be held against the bottom wall of the mold by hand, generally using forceps. If this is not the case, it must have the ability to appropriately cut out the tissue later on with a microtome. The molten paraffin is cooled appropriately on a refrigerated plate (which may be a thermoelectric cooler (TEC)) in order to partially solidify the paraffin so that it is suitable against the bottom wall of the mold. Hold the tissue specimen in the orientation. The cassette is then placed on top of the base mold and paraffin is injected into the base mold through the open top of the cassette. The cassette changes its function at this point in the procedure from a component holding tissue to a fixation device for later use in cutting or cutting the solidified paraffin in a microtome. The base mold is cooled until all the molten paraffin has solidified and the histologist has removed the stainless steel base mold from the embedded paraffin block. Thus, the tissue specimen is used as a holder in a microtome chuck and is embedded in a paraffin rectangular block with a plastic tissue cassette on opposite sides. As with the tissue processing device, the embedding process is accomplished in a batch format. During this time, the average histologist can embed approximately 40-60 cassettes per hour.

  The solidified paraffin block comprising the embedded tissue specimen is then ready to be cut into ultrathin sections for mounting on a microscope slide. The histologist places the embedded tissue block in a chuck on a microtome that is sized to accept the side of the block with an embedded plastic cassette. The histologist then begins to cut the paraffin block having the tissue specimen embedded on the opposite side of the surface of the plastic cassette. Individual ribbon-like tissue slices embedded in the paraffin are obtained. When the microstorm operates normally, the individual slices are brought together. These very thin ribbon-like slices are then floated in a water bath and a glass slide is placed under the slices. Then, the slice in which the thinly cut tissue specimen is embedded is attached to the upper surface of the slide.

  If the histologist obtains a sufficient number of slides from the tissue specimen, the slides are placed in an automatic staining apparatus. The staining device stains different tissues and cells of the slide in different colors through a series of infiltration stages. This allows pathologists to recognize different structures and more easily find abnormalities in the tissue. After the staining procedure is complete, the slide is covered with a coverslip and the pathologist is placed under a microscope and ready for analysis. Based on the above outlined procedure, it is understood that traditional tissue specimen management and processing involves a number of manual steps performed by histologists and is a very labor intensive process. Should be. For example, repetitive pressure disorders such as carpal tunnel syndrome are prevalent. This is especially true for the process of embedding tissue specimens. These numerous manual processes and repetitive management of tissues increase the possibility of human error, and the tissue specimens finally attached to slides for analysis by pathologists are ideal for accurate diagnosis In order to ensure proper condition and orientation, highly trained and skilled histologists are required.

U.S. Patent No. 6,028,056 (the '032 patent) discloses various improvements to the technical field to which the present invention belongs, including a new method of holding tissue specimens during gross-in, embedding, and microtome or excision procedures. . More specifically, Patent Document 1 relates to a device that is a cassette and can be cut out with a microtome, and confines and supports tissue. When a cassette is used, the tissue specimen is fixed in the cassette and relies on a process for exchanging tissue fluid for paraffin. Next, since the tissue specimen and the cassette are placed on a microscope slide, they are cut out simultaneously. Since the tissue specimen is never removed from the cassette from when it is processed by the tissue processing apparatus to when it is cut out by a microtome, management time can be saved to a significant degree. Further, the loss of the organization, for example due to human error or dropping the organization during management, is greatly reduced by eliminating the individual organization management steps. The patent also schematically discusses an automated process that, in combination with a new tissue cassette, further reduces the management steps between the procedures.
US Pat. No. 5,817,032

  Despite various advances in the technical field to which the present invention pertains, slices or ribbons that manage tissue specimen procedures, improve manufacturing capabilities and quality in embedded tissue specimens, and are susceptible to diagnosis There is a growing need for further improvements in methods that facilitate the creation of tissue embedded tissue.

  The present invention generally provides a cassette for holding tissue specimens. The cassette includes a body and may further include a lid. The body includes a bottom wall and at least one side wall that extends upwardly relative to the bottom wall to form an interior space for receiving the tissue specimen. It comprises one or more sides. In one aspect, a plurality of query points (eg, sensor elements) are associated with the body and / or lid, and an automated sensing system is configured to determine at least one characteristic of the cassette. ing. The characteristic refers to size, shape, or some other structural or functional characteristic. The cassette preferably includes a lid that is coupled to the body and configured to be movable between an open position and a closed position. A flange extends along the top of the at least two side walls, and the sensor element is provided on the flange. The sensor element further comprises an opening or other detectable element and a computer readable property or element. The detection can be performed by a contact type or a non-contact type. Aspects of the present invention may also be applied to, or alternatively, a frame member that holds the cassette during processing and / or embedding procedures. The bottom wall of the cassette body preferably transitions to a plurality of side surfaces having rounded corners. The bottom wall is radially elongated and / or wide in the direction from the gate or fill location near the cassette body in the mold used to manufacture the cassette to a plurality of sides. It is preferable to further include a plurality of openings. Each of these features can help guide the material flow to form the cassette during the injection molding process. It is preferable that the opening of the bottom wall is provided at substantially the center of the bottom wall and has a teardrop shape and / or an oval shape.

  At least two side surfaces are positioned at positions facing each other, and a distance between the facing side surfaces varies along a longitudinal direction of the facing side surfaces. As a result, the side surface can be bent at a certain angle along the longitudinal direction, and can be undulated or rounded, which helps to cut out more easily with a microtome. By way of example, the side surfaces can be bent at an angle from one end to the opposite end, or from a position between the opposite ends to each opposite end.

  In another aspect, the stopper member in the internal space of the cassette is configured to stop the lid at a minimum distance from the upper surface of the bottom wall. The stopper member may be a part of the cassette body or a part of the lid.

  In another embodiment, the orientation cassette comprises a plurality of posts extending upward from the bottom wall. The post is configured to determine the orientation of the tissue specimen therebetween to ensure that the section taken as the specimen is most appropriate for diagnostic purposes. The lid that is receivable in the interior space includes an opening for removably receiving the post. The posts are arranged so as not to align with each other on a straight line that is perpendicular or parallel to any one of the side surfaces. This helps to ensure a more effective cutting with the microtome and less fuzzing of the cutting edge of the microtome.

  Upon review of the following detailed description of the preferred embodiment in conjunction with the drawings, those having ordinary skill in the art to which the present invention pertains will immediately be able to understand these and other objects, advantages and features.

[Detection of cassette structure / shape]
With reference to FIGS. 1 and 2, the cassette 10 is utilized in an automatic embedding device. For example, the apparatus is disclosed in the above-mentioned incorporated WO 04/029584. More specifically, the apparatus can utilize a photoelectric identification function that determines which of the at least two types of cassette 10 is removed from the input basket by a robotic system. For example, different types of cassettes 10 can be of different sizes or have other different characteristics and / or functions. A plurality of query points, which are openings 12 provided in the flange 14 at the outer peripheral portion of the cassette body 16, are transmitted to the optical reader by a binary signal at each query point on the cassette 10. It can be used to grant. At this time, since it is advantageous that all cassettes 10 use the same embedding frame, all the cassettes 10 have substantially the same alignment accuracy at the upper side position and the lower side position. Designed to engage the inside of the frame. If additional functions of the cassette are developed or customer identification is required, for example, similar sensor functions are incorporated into the frame that receives the cassette 10. The function is incorporated in many different types of cassettes and used in combination with automated machinery.

  In the co-pending application WO 04/029584, the sensor queries each cassette 10 after it is removed from the input basket. The robot arm moves the cassette 10 in the frame below the single sensor head. The sensor detects the presence of the cassette to ascertain whether the cassette 10 has dropped from the input basket or is improperly gripped. More specifically, the sensor detects a binary signal (signal vs. no signal) in cooperation with the machine control unit in order to shift to the embedding process of the cassette 10. The function can be extended so that the sensor and control device transitions to determining how to handle the cassette 10 based on the size, type and / or other parameters of the cassette. Other parameters may include the base mold type and any special heating or cooling parameters required to process and / or embed a particular type of cassette. The sensors are used, for example, to inspect each cassette assembly / frame assembly at each input position of the plurality of positions programmed to inspect the signal by the controller. As described above, the four openings 12 at the corners of the flange 14 are currently utilized for the input position. The central portions 14a and 14b of the cassette flange 14 are used to increase the number of cassettes that can be identified and implemented by the machine. As will be described later, by providing the opening 12 at the corner, three types of cassette configurations are discovered. Further, since the cassette 10 has symmetry and is attached by an operator in two different orientations inside a frame (not shown) for carrying the cassette 10 through the process, the opening 12 is provided in the corner portion. That includes redundant routines. The automatic machine must accurately detect and inspect the cassette configuration with high accuracy. Using a redundant detection area is one method for easily realizing the improvement of certainty.

  The sensor in a preferred embodiment is an emitter / detector sensor in which the light to be detected (infrared or other color spectrum) is directed to the tuning sensor. If the query point is open (e.g., an opening 12 in the cassette flange 14 is detected), the emitter receives a signal to the resistor sufficiently. If the query point is opaque or diffuse (eg, the opening 12 is not detected), the tuning sensor cannot receive enough light to record a signal. Since the cassette assembly is pre-processed in a tissue processing apparatus comprising utilizing a solution containing paraffin, a paraffin concavo-convex lens that obstructs one of the sensor openings 12 is provided. There is a possibility of being created. The sensor opening 12 can be made large enough to minimize the possibility.

  Furthermore, the cassette 10 has a rectangular shape having two long sides and two short sides, and as described above, along the longitudinal axis of the cassette 10. Can be mounted in the frame in two different orientations. The configuration of the detection unit is configured in consideration of this. In other words, the configuration allows the machine to identify the three cassette types regardless of the orientation of the cassette 10 in the frame. Each longitudinal side provides sufficient information to properly detect the cassette type, and the other longitudinal side carries redundant information. If more than two configurations are required, then a sensing hole (not shown) may be additionally provided on either side of the center line of the end.

  According to the actual detection plan, three possible cassettes are as follows:

[Cassettes for large tissues]
All corners are opaque (there is no opening 12).
Signal: Both sensors are off.

[Biopsy cassette (small cassette) 10]
The opposite corner is opaque (there is no opening 12). The other corner is opened (the opening 12 exists). Symmetrical with respect to the diagonal.
Signal: One sensor is on, but another sensor is off (does not respond to commands).

[Orientation cassette]
All corners are opened (opening 12).
Signal: Both sensors are on.

  It should be understood that many different types of sensor systems (including either contact type sensors or non-contact type sensors) can be used to implement the technical idea of the present invention described above. . For example, many different types of optical sensor systems, magnetic sensor systems, barcode-type systems or RFID-type systems can be used with the cassettes and / or frames read out under the control of automated embedding systems and / or processing systems. It can be used to properly identify other forms of information. As one additional example, the frame member 90 is encased therein for properly embedded information to be relayed to the control system (eg, automated processing and / or embedding device). The inclusion of a buried RFID element 81 is shown in FIG. The communicated information is also extensive and includes, for example, various diagnostic information, patient history, tissue specimen information or information useful for histological or pathological processes.

[Fixing the cassette in the frame member]
According to FIGS. 1 and 8, a plurality of depressions 80, 82 and 84, 86 are formed on the upper surface of the flange 14 along the side surface along the longitudinal direction of the cassette body 16. Frame member 90 (shown in FIG. 8) is utilized to carry cassette 10 and recesses 80, 82 and 84, 86 receive protrusions or tabs 102, 104 and 106, 108, respectively. The recesses 110, 112 and 114, 116 can be additionally formed on the upper surface of the flange 14 along the side surface of the cassette body 16 along the short direction. The recesses receive protrusions or tabs 120, 122 and 124, 126 formed in the frame member 90, respectively. The registered tabs and recesses securely hold the cassette 10 within the frame member 90 during processing and embedding operations.

[Air removal during embedding process]
The tissue embedding process, when performed manually using conventional cassettes, requires steps or techniques that eliminate or reduce air bubble confinement within the block of paraffin (or embedding material). The bubbles do not have a positive effect because the bubbles make the paraffin block more fragile, and then the microtome cutting "breaks out" or breaks the main paraffin body. This must remove inconsistent edges or openings in the “ribbon” -like piece and flow continuously from the piece in the microtome process. This can result in failed ribbons and poor quality diagnostic slides. One way to reduce the presence of air bubbles is to tap the cassette assembly to remove the air bubbles, otherwise move it quickly. However, the machine is configured to perform such steps by an automated, high-speed embedding process. Thus, where air accumulates, the largest possible opening is provided that the air can be eliminated during the paraffin filling stage. 1 and 2, openings 12, 20 in the flange 14 of the cassette 10 and below the writing surface of the frame are provided that air can be eliminated. Accordingly, it should be understood that the sensor opening 12 has two purposes. These openings 12, 20 are large enough that air is excluded and there are no burrs or protrusions. An obstacle such as a rib under the flange suppresses the generation of bubbles. A reinforced flange 22 is provided on the upper surface of the flange 14 to eliminate air trapping. The frame member 90 also includes an air escape recess or passage 83 on its inner surface.

[Material flow to injection molding cassette]
Referring again to FIGS. 1 and 2, certain functions of the cassette 10 prepare for an effective injection molding technique to be utilized during its manufacture. First, the gate for filling the cassette body 16 and lid 24 mold has a flow runner that leads directly to the end of the cassette side wall 26. At the bottom wall 28, it is most desirable for the cassette body 16 to prevent the tissue from having an area that is obstructed from the flow path used to process it. Although the center line 28a of the bottom wall 28 has several preventable gates, most of the area is covered by the inflow opening 30. For molding, it is generally preferred to have a solid flow rib 24a as used for the lid 24. In an aspect of the present invention, a mold pin for forming a central opening (shown in FIG. 1B) is configured with a teardrop shape to direct the flow of material of the cassette outward toward the side wall 26. Has been. Another mold pin for forming the opening 30b is formed radially and elongated toward the side wall 26. These shapes help to direct the cassette material away from the gate or filling position (eg, central region 28a). The transition region 29 (shown in FIG. 3) from the bottom wall 28 to the rib 40 forming the side wall 26 is radiused to reduce the inflow restriction for molding.

[Non-linear side wall]
1 and 2 also show that the side walls 26 are non-linear with respect to the frame side which also determines the cut surface of the microtome. As shown in the figure, this means that the side surface 26 is bent at a substantially steep angle from its intermediate point 26a, or the distance between the opposing side walls 26 depends on the angle of the side wall 26 in the longitudinal direction or the degree of bending. It means to change. For example, the side 26 of the cassette has a large radius along its length, or is wavy on the side of the frame. This feature aims to prevent having a side wall parallel to the side surface of the frame in order to improve the cutting efficiency of the microtome. It should be understood that many configurations can accomplish this purpose.

[Minimum lid engagement height]
According to FIGS. 1A, 3 and 4, the lid 24 is a side wall bump or protrusion 50a, 50b or 50c to maintain engagement with the cassette body 16 at an appropriate height relative to its tissue specimen. Is engaged. This encloses and secures the tissue specimen in cassette 10 during processing. Even extremely small tissue specimens such as biopsy specimens with a size of about 1 cubic millimeter can be included and fixed. The lid 24 does not fall all the way and does not break sensitive biopsy specimens. The stopper 52 is attached to the cassette body 16 so that the lid 24 does not fall on the specimen during processing. Such a stopper can alternatively be provided on the lid 24. The height of each stopper 52 is about 0.75 mm, but the height may be in the range of about 0.25 to 1 mm. Also, the bottom wall 28 (preferably about 0.38 mm) of the ultra-thin cassette allows the specimen to be confined, but detrimental damage revealed by the cut biopsy and subsequent diagnostic slides. It does not have undue compressive force that causes detrimental crush artifacts.

[Biopsy material orientation cassette]
According to FIGS. 6 and 7, the orientation cassette 60 is used to determine the orientation of a special tissue (not shown) that must have been cut “on edge”. Corresponding to the reference numerals of the first embodiment, the reference numerals having the symbols first appearing in FIGS. 6 and 7 refer to the corresponding elements of the structure. Tissues such as skin, gal bladder, bladder, etc. need to be oriented so that the pathologist can see a complete section of the biopsy material. These specimens are extremely small. Therefore, the specimen needs to be held in a cassette having a small opening, such as the biopsy cassette 10. The biopsy material orientation cassette 60 allows a pathologist to place the tissue in an appropriate cut-out direction during gross-in. This predetermined orientation is maintained through a procedure for processing and embedding the tissue, so that it is not necessary to determine the orientation again in order to remove the tissue and cut out the paraffin mold. Is maintained. The tissue is placed between upright posts 62 formed on the bottom wall 64 of the cassette 60. These posts 62 are arranged in a configuration that takes into account the average thickness and length of the tissue utilized in the cassette 60. The posts 62 are further arranged so that they are not directly aligned with each other either parallel or perpendicular to the cutting blade path. The post contains more plastic material than the side ribs 66, which causes the microtome blade to blunt at an undesirable rate after repeated use. Thus, the posts 62 are positioned to minimize the number of posts 62 that are continuously cut out in the same blade path. The lid 70 of the orientation cassette 60 has a corresponding clearance hole 72 for the upright post 62. Like any other tissue confining the cassette configuration disclosed herein, or an incorporated application product, the tissue is held in place between the posts 62 and the lid 70 sinks over the posts 62. To fix the tissue so that it does not move. There is a small gap between the lid relief groove 70 and the post 62 to prevent any tissue from exiting the cassette 60.

  While the invention has been described in terms of various embodiments thereof, the embodiments have been described in considerable detail and are not intended to limit or limit the scope of the claims to such details. . Apart from and / or in addition to the detailed examples and principles disclosed herein, the components disclosed herein may be improved with the detailed examples or principles described in the above-incorporated PCT application. Can be done. Additional advantages and improvements will be readily apparent to those skilled in the art to which the present invention pertains. Accordingly, the invention in its broader aspects is not limited to the specific details, representative apparatus, techniques, and illustrative examples described and represented. Accordingly, departures not described herein may be created from such details without departing from the scope of the invention or the applicant's general inventive concept. The various features of the present invention may be considered as being independent or not as disclosed herein, whether or not such combinations are explicitly disclosed as examples shown and described in the detailed description of the invention. It can be used in various combinations.

The perspective top schematic diagram of the cassette for biopsy comprised in this invention is shown. The enlarged view of 1A part enclosed with the circle | round | yen of FIG. 1 is shown. FIG. 2 shows an enlarged top view of the bottom wall of the cassette represented in FIG. 1. FIG. 2 shows a perspective bottom wall view of the biopsy cassette shown in FIG. 1. Sectional drawing in the cross section 3-3 of FIG. 1 is shown. Sectional drawing in the cross section 4-4 of FIG. 1 is shown. Sectional drawing in the cross section 5-5 of FIG. 1 is shown. The perspective top schematic diagram of the tissue orientation cassette constructed in the present invention is shown. FIG. 7 shows a top view of the tissue orientation cassette shown in FIG. 6. FIG. 4 shows a perspective view of a frame member constructed and adapted to carry a cassette in the present invention.

Explanation of symbols

DESCRIPTION OF SYMBOLS 10 Cassette 12 Opening part 14 Flange 16 Main body 20 Opening part 24 Cover 26 Side wall 28 Bottom wall 60 Cassette 62 Post

Claims (6)

A cassette that can receive and embed a tissue specimen, and then cut out the tissue specimen with a microtome,
Includes a bottom wall and at least one sidewall, said sidewall to form an interior space for receiving the tissue sample was stretched upwardly relative to the bottom wall is coupled to frame and removable A body configured as follows,
Provided to the body, a plurality of query points that are configured to at least one characteristic of the cassette is determined by the automatic sensor system,
A cassette for holding a tissue specimen characterized by comprising:   The lid according to claim 1, further comprising a lid that is configured to be coupled to the main body, is movable between an open position and a closed position, and has higher rigidity than the bottom wall. Cassette.   The cassette of claim 1, further comprising a flange extending along an upper portion of the side wall and comprising the query point.   The cassette of claim 3, wherein the query point further comprises an opening.   The cassette of claim 1, wherein the query point further comprises an opening. A cassette that can receive and embed a tissue specimen, and then cut out the tissue specimen with a microtome,
A bottom wall and a body comprising at least one side wall, said side wall, in order to form an interior space for receiving the tissue sample, the stretched upwardly relative to the bottom wall, frame and removable A body configured to be coupled, and
A lid configured to be coupled to the body;
At least one in formed among the body and the lid, and a plurality of query points that are configured to at least one characteristic of the cassette is determined by an automated sensor system,
Cassette for holding a tissue specimen, characterized in that it comprises a.

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