Apparatus and method for transporting tissue sections

Abstract

The present invention relates to an apparatus and method for automated tissue sectioning, including slicing and transferring tissue sections to slides.The present invention also relates to an automated system and method for transferring tissue sections onto slides using adhesive strips.

Description

Technical Field

[0001] The present invention relates to an apparatus and method for transferring tissue sections from a microtome to slides, including an automated system and method.

[0002] [Cross - Reference to Related Applications] This application claims the priority and benefit of U.S. Patent Application No. 63 / 147,392, filed on February 9, 2021, the content of which is incorporated herein by reference in its entirety.

Background Art

[0003] A microtome is used to cut extremely thin slices of tissue, known as tissue sections, for microscopic examination under transmitted light or electron radiation. With a microtome, a very sharp knife is used to slice thin tissue sections of a desired thickness from a sample. The knife is held by a knife holder that can be adjusted to cut at a selected thickness. Thereafter, the tissue sections are typically recovered from the microtome by manual retrieval, such as by the operator using a gripping tool, transferred to a slide, and can be examined using an optical or electron microscope or by other techniques. Since tissue sections are extremely thin, some having sub - micron thicknesses, it can be difficult to collect and transfer them without damage or contamination.

[0004] Rotary microtomes, such as the Leica RM2125 RTS, include a handwheel that moves a sample holder downward so that a held sample is sliced by a knife. Rotary microtomes have a housing in which a knife holder, a sample holder, a handwheel and other elements are all assembled and have various clamps for holding moving parts in place. The knife holder typically includes a front plate, and sliced tissue sections can be placed on the front plate after slicing. Slicing is a manual operation, and each section is sliced by rotation of the handwheel.

[0005] Patent Document 1 by Thiem et al. discloses a rotary microtome having a base portion on which a microtome housing and a knife holder are arranged. The collection element surrounds the knife holder on three sides and has a U-shaped base outline. Several clamp levers are associated with the knife and knife holder, and the clamp levers for the knife holder are located outside the collection element, reducing the number of levers for the knife holder and simplifying cleaning. The collection element is configured to allow for easy removal of tissue sections and to prevent the user from becoming entangled in the collection element or the microtome housing.

[0006] Patent document 2 by Walter et al. discloses a device for applying histological sections to slides. Histological sections are produced by the cutting action performed by the blades of a microtome. The device comprises a positioning device having a component that is rotatably mounted on a bearing and has a receptacle that receives and holds the slide, the positioning device being designed to rotate the slide received in the receptacle around the axis of rotation of the rotatably mounted component.

[0007] Patent document 3 by Schmitt discloses an apparatus and method for separating histological sections produced using a microtome. Previously produced histological sections and currently produced histological sections are connected to form a section strip. To simplify the separation of the previously produced and currently produced histological sections from each other, a nozzle device is provided, which causes the histological section positioned on the blade holder to receive a directional and intensity-controlled airflow such that the histological sections of the previously produced section strip are separated from the currently produced histological section positioned on the blade holder and removed from the blade holder.

[0008] Patent document 4 by Magavi et al. discloses an apparatus and method for handling a portion of a tissue sample sectioned by a microtome. The apparatus comprises a container, a tissue sample holder within the container, and one or more outlets configured to allow fluid to flow from the container. The flow through the outlets moves the portion of the tissue sample sectioned by the microtome toward the outlets. The method includes sectioning one or more portions of a tissue sample and flowing fluid through the tissue sample to move one or more portions of the tissue sample toward at least one fluid outlet.

[0009] Patent document 5 by Williamson et al. discloses an automated machine for handling and embedding tissue samples contained on sectionable supports of a microtome. The machine comprises an input member configured to hold multiple sectionable supports of a microtome before a tissue embedding operation. An output member configured to hold multiple sectionable supports of a microtome after a tissue embedding operation. A cooling unit is configured to hold at least one of the sectionable supports of the microtome during a tissue embedding operation. A motorized carrier assembly is mounted to move and configure to hold at least one of the sectionable supports of the microtome. The carrier assembly moves the support from the input member to the cooling unit and finally to the output member. A feeding device feeds the embedding material onto the sectionable supports of the microtome and at least one tissue sample supported by the sectionable supports of the microtome during the embedding operation.

[0010] Traditional methods of slicing tissue sections using a microtome involve creating strips or ribbons of tissue sections from a tissue sample (such as tissue embedded in paraffin). After the tissue sections are prepared, the posterior edges of the sections are adhered to the knife, allowing for adhesion between the sliced ​​tissue sections and those to be sliced ​​later, thus creating a ribbon of tissue sections. Because it is difficult to manipulate the first section sliced ​​by the microtome without damaging it, in practice, only one or two sections from the ribbon are often used and mounted on the slide. This results in the first section becoming a sacrificial section, wasting sample. The first section of the ribbon often curls or collides with the knife holder, requiring the microtome user to intervene to remove the tissue section. If the first section sliced ​​from the sample block can be obtained without this user intervention, the first section can be used directly, eliminating the need to create a ribbon.

[0011] Patent document 6 by Studer discusses a device for transferring section ribbons to a specimen holder for use in a transmission electron microscope (TEM), in which the section ribbons are transferred to the specimen holder by an ionization device within a microtome apparatus.

[0012] Patent document 7 by Orfield et al. discusses the need to improve systems for mounting tissue sections onto slides. Generally, in biological laboratories, it is desirable to mount tissue sections onto slides for the purpose of examining them using a microscope, processing them with colorants or dyes, and for other purposes. Conventional systems and methods for mounting specimens onto slides involve placing the tissue sections in a water tank of sufficient depth, with the specimens floating on the water surface. The narrow side of the slide is then rested on the edge of the water tank, and the slide is angled downward into the water so that it is partially submerged. The tissue sections are then manipulated onto the slide using a small brush or glass capillary. Typically, the slide is gradually withdrawn from the water as additional tissue sections are placed on the slide. In another modification of the conventional method, the tissue is embedded in paraffin wax, sliced ​​using a microtome, and then selected sections of the embedded tissue are transferred to a heated water tank. The hot water bath partially melts the paraffin around the specimen, and then the tissue section is scooped out of the hot water bath using a glass slide treated with an adhesive. Therefore, conventional methods of mounting specimens onto slides are difficult, time-consuming, and labor-intensive.

[0013] Orfield further discusses a system and method for mounting sections onto a substrate. This system comprises a fluid channel having a fluid channel inlet that receives sections processed from a bulk-embedded sample by a sample sectioning module positioned near the fluid channel inlet, a section mounting region downstream of the fluid channel inlet, and a fluid channel outlet downstream of the section mounting region, a reservoir fluid-communicated with the fluid channel outlet, and a manifold fluid-coupled to the reservoir and transmitting a fluid flow that drives the delivery of sections from the fluid channel inlet toward the section mounting region by delivering fluid from the reservoir to the fluid channel inlet.

[0014] Patent document 8 by Angros discusses a method for removing a suspension from between a microscope slide and a paraffin-embedded biological specimen. The microscope slide on which the paraffin-embedded biological specimen is suspended is positioned on a slide support element. As the slide support element rotates, the microscope slide and the paraffin-embedded biological specimen move in such a way that the suspension that is positioned between the microscope slide and the paraffin-embedded biological specimen is removed from between them.

[0015] Lim et al.'s Patent Document 9 discusses an apparatus and method for automated tissue sectioning, including slicing and transferring tissue sections from a microtome. This published patent also discusses an automated system and method for slicing and transferring tissue sections onto slides.

[0016] Lim's Patent Document 10 discusses a method and apparatus for transferring tissue sections from a first location to a second location. A thermoplastic material is applied to at least one pin of the tissue section transfer apparatus. The thermoplastic material comes into contact with the tissue section while at least partially melting, and when cooled, the substantially solid thermoplastic material holds the tissue section to at least one pin. The tissue section transfer apparatus comprises a heater block, a heat source, and at least one pin attached to and extending from the heater block.

[0017] Patent document 11 by Ornstein describes a method for transferring thin tissue sections, attached to a substrate by a first pressure-sensitive adhesive layer, to a microscope slide supporting a second pressure-sensitive adhesive layer made of a polymerizable material. The tissue sections come into contact with the second adhesive layer to form a laminate. After the second layer is polymerized to form a secure bond between the tissue sections and the microscope slide, the substrate and the first adhesive layer are completely removed, exposing the tissue sections currently attached to the microscope slide by the polymerized second adhesive layer for subsequent processing. [Prior art documents] [Patent Documents]

[0018]

Patent Document 1

Patent Document 2

Patent Document 3

Patent Document 4

Patent Document 5

Patent Document 6

Patent Document 7

Patent Document 8

Patent Document 9

Patent Document 10

Patent Document 11

Summary of the Invention

Problems to be Solved by the Invention

[0019] An apparatus and method for automatically collecting tissue sections from a microtome and transferring them to a slide, particularly an apparatus and method that increase speed and efficiency while maintaining the high quality of the tissue sections, are desirable.

Means for Solving the Problems

[0020] As one aspect of the present invention, a method for transferring a tissue section onto a slide is provided. The method includes collecting the tissue section using a tissue collector and / or an adhesive strip (i.e., a tissue collector or an adhesive strip or both). The method also includes moving the tissue collector and / or the adhesive strip to bring at least a portion of the tissue section into contact with the slide, and applying a physical force, heat, or both to the tissue section at the contact location between the tissue section and the slide. Using this method, the tissue section can be easily and reliably transferred onto the slide without manual contact with the tissue section.

[0021] The method can include positioning the adhesive strip on the collection end of the tissue collector, and the tissue section can be collected by adhering it to the adhesive strip. In some embodiments, the adhesive strip is provided or wound on a first reel, and the adhesive strip is extended or unwound from the first reel across the collection end to a second reel. The adhesive strip can be advanced across the collection end (e.g., by unwinding a portion from the first reel and winding a portion onto the second reel) after the first section of the adhesive strip has been used to transfer the tissue section onto the slide. In this way, the second unused section of the adhesive strip can be positioned on the collection end.

[0022] In another embodiment, a device for transferring tissue sections to a slide is provided. The device comprises a tissue collector configured to collect tissue sections, a slide holder configured to hold the slide, and a heating element. The tissue collector and / or slide holder (i.e., the tissue collector or the slide holder or both) are configured to move so that the tissue sections collected by the tissue collector and the slide held by the slide holder come into contact at a contact point. The heating element is configured to be positioned at the contact point or to move at the contact point. The tissue collector and / or slide holder can be configured to perform rotational, linear, or both movements. In some embodiments, the tissue collector and / or slide holder are configured to move automatically.

[0023] In some embodiments, the heating element is housed within a tissue collector or slide holder. Additionally or alternatively, the heating element is located outside the tissue section and slide holder, and is positioned on or movable to a first surface of the slide held by the slide holder, with the contact point being a second opposing surface of the slide. The heating element can be configured and positioned to provide heat to the top of the tissue section. In some embodiments, the device further comprises an adhesive material dispenser.

[0024] In yet another embodiment, a device is provided for transferring tissue sections to a slide, comprising a tissue collector for collecting tissue sections, a slide holder for holding the slide, and an adhesive strip dispenser capable of supplying an adhesive strip at the collection end of the tissue collector. The tissue collector and / or slide holder may be configured to move such that the tissue sections collected by the tissue collector and the slide held by the slide holder come into contact at the contact point. In some embodiments, the adhesive strip dispenser comprises at least two reels on either side of the collection end, with the adhesive strip extending from a first reel across the collection end to a second reel. The first reel is capable of unwinding a new adhesive strip, and the second reel is capable of winding a used adhesive strip. Since it is advantageous for the collection end of the tissue collector to be extendable to facilitate the collection of tissue sections, the adhesive strip dispenser should be configured not to interfere with or prevent the extension of the collection end. In some embodiments, the tissue collector is configured to transfer tissue sections by applying physical force and / or heat (i.e., physical force or heat or both) to or near the slide holder. In some embodiments, the tissue collector or slide holder includes a heating element.

[0025] The above apparatus may further include a microtome comprising a sample holder, a knife holder, and a knife held by the knife holder and facing the sample holder, wherein when the sample holder moves linearly, the sample held by the sample holder is sliced ​​by the knife to form tissue sections. The above apparatus may also include a slide chamber, and the slide holder may be configured to immerse the slides in the slide chamber.

[0026] The method may further include slicing tissue sections from a tissue sample using a microtome, extending a tissue collector to collect tissue sections from the microtome, and / or adhering the sliced ​​tissue sections to the tissue collector using adhesive, reduced pressure, friction, or other force (i.e., slicing tissue sections from a tissue sample using a microtome, extending a tissue collector to collect tissue sections from the microtome, or adhering the sliced ​​tissue sections to the tissue collector using adhesive, reduced pressure, friction, or other force, or all of these). The method and apparatus may further include moving the slide holder and the adhering tissue sections into a slide tank containing liquid, and / or immersing the slide and the adhering tissue sections in liquid (i.e., moving the slide holder and the adhering tissue sections into a slide tank containing liquid, or immersing the slide and the adhering tissue sections in liquid, or both).

[0027] The above apparatus and method may also include a control system configured to automate the heating of tissue sections by a heater. The above apparatus and method may also include a user interface that communicates with a controller, the user interface being configured to display or report the availability of the adhesive material. [Brief explanation of the drawing]

[0028] [Figure 1] This figure shows an exemplary embodiment of a tissue collector equipped with a heating element. [Figure 2] This figure shows an exemplary embodiment of a slide holder equipped with a heating element. [Figure 3] This is a perspective view of one exemplary embodiment of a device that automatically slices tissue sections and transfers the tissue sections to a slide. [Figure 4] This is a diagram of an exemplary embodiment of a tissue collector for holding tissue sections, which includes an adhesive strip dispenser. [Modes for carrying out the invention]

[0029] Some embodiments of the present invention are at least in part based on reducing or eliminating the manual step of collecting tissue sections from a microtome and transferring them to a slide. Some embodiments of the present invention are also based on an improved method for mounting tissue sections to a slide. Tissue sections are very thin slices of tissue that are suitable for microscopic examination, for example, slices having a thickness of about 1 μm to about 100 μm, alternatively about 1 μm to about 30 μm, or alternatively about 3 μm to about 10 μm. Manually transferring tissue sections to a slide requires care and skill.

[0030] In some embodiments of this method and apparatus, tissue sections are attached to a slide by applying heat, physical force, or both to the contact point between the tissue section and the slide. Heat, physical force, or both can be applied by a tissue collector and / or slide holder (tissue collector or slide holder or both). The tissue collector collects tissue sections from a microtome used to slice tissue sections from a tissue sample. The tissue collector may collect tissue sections from other devices or locations other than a microtome, for example, from a storage location. The tissue collector may be configured to apply heat, physical force, or both to the tissue section after it has been moved and brought into contact with the slide. The slide may be held by a slide holder, and in some embodiments, a heating element located on or within the slide holder can apply heat to the tissue section. The tissue collector and slide holder may be part of a larger apparatus that results in the automation of the tissue collector and / or slide holder.

[0031] In some embodiments, the tissue section is heated at the contact site to an arbitrary temperature between approximately 40°C and approximately 90°C, or a temperature sufficient to at least partially melt the embedding medium. Alternatively or additionally, a physical force is applied to the tissue section, such as by applying an arbitrary physical force of approximately 275 kPA or less, for example, approximately 200 kPA or less or approximately 125 kPA or less in some embodiments, to the contact site of the tissue section. In some embodiments, the physical force is at least 5 kPA or at least 25 kPA. In some embodiments, no physical force is applied to the contact site of the tissue section. If the tissue section contains an embedding medium such as paraffin, it may be desirable that the heat is sufficient to at least partially melt the embedding medium, and / or that the physical force is sufficient to deform or compress the embedding medium (i.e., the heat is sufficient to at least partially melt the embedding medium, or the physical force is sufficient to deform or compress the embedding medium, or both). In some embodiments, the embedding medium is at least partially melted and solidified at the contact site and / or at least partially compressed at the contact site (i.e., released by at least partially melted and solidified at the contact site, or at least partially compressed at the contact site, or both).

[0032] The contact area can be an area of ​​any size, shape, or other configuration. For example, the contact area can be a thin, straight portion of the tissue section at or near the apex, extending from one side of the tissue section to the other, or across the width of the tissue section. In some embodiments, the tissue section can be considered to have an apex, a base, and a side extending between the apex and base, which can be particularly useful when the apex of the tissue section is held and the rest extends downward due to gravity. In some embodiments, the tissue section or a portion thereof is attached to the contact area of ​​the slide by, for example, melting the embedding medium on the slide. In this situation, the tissue section is attached if it remains on the slide as the tissue section and slide move into the tissue chamber. This attachment does not need to be permanent or particularly strong. When the slide is vertical, or when gravity separates the tissue section from the slide, it is generally sufficient if the attachment overcomes gravity.

[0033] In some embodiments, the contact area and / or attachment area (i.e., the contact area and / or attachment area or both) is located at or near the apex of the tissue section. The contact area and / or attachment area is located at the apex if it includes the end or margin of the tissue section, and near the apex if the end or margin is outside the contact area. For example, a linear contact area is near the apex when it is 1 mm or 2 mm below the apical margin. The contact area can be selected or predetermined to be smaller than the entire area of ​​the tissue section, or to include only the embedding medium and no tissue. In some embodiments, no physical force or heat is applied to the tissue section outside the contact area. In some embodiments, the contact area and / or attachment area includes an area of ​​50% or less, or 40% or less, or 25% or less, or 20% or less, or 15% or less, or 10% or less, or 7.5% or less, or 5% or less, or 2.5% or less, or 1% or less of the tissue section.

[0034] In some embodiments, the method may further include slicing tissue sections from a tissue sample using a microtome, extending a tissue collector to collect tissue sections from the microtome, and / or adhering the sliced ​​tissue sections to the tissue collector using adhesive, reduced pressure, friction, or other force (i.e., slicing tissue sections from a tissue sample using a microtome, extending a tissue collector to collect tissue sections from the microtome, or adhering the sliced ​​tissue sections to the tissue collector using adhesive, reduced pressure, friction, or other force, or all of these). In some embodiments, the method may further include moving the slide holder and the adhering tissue sections into a slide tank containing a liquid, and / or immersing the slide and the adhering tissue sections in a liquid (i.e., moving the slide holder and the adhering tissue sections into a slide tank containing a liquid, or immersing the slide and the adhering tissue sections in a liquid, or both). Other steps and uses will be apparent from the following description of the method and apparatus.

[0035] Figure 1 shows an exemplary embodiment in which a tissue collector 102 holds the apex of a tissue section 104. The tissue section 104 is substantially vertically oriented and is mostly free or unrestricted, loosely suspended from the apex 108 held by the tissue collector 102. The tissue section 104 contains tissue 110 in an embedding medium 112 such as paraffin. The tissue collector 102 is movable and extends in a substantially horizontal motion toward the slide 106 in the arrangement or orientation shown in Figure 1. In some embodiments, the tissue collector 102 attaches the tissue section 104 to the slide 106 by applying a physical force to the tissue section 104 on the contact slide 106. For example, a physical force of about 275 kPA or less, or any amount of physical force sufficient to cause attachment without damaging the slide, can be applied. The tissue section 104 can be attached to the slide 106 as a result of deformation, compression, or other mechanical action. As an alternative to, or in addition to, the application of physical force, the tissue collector 102 may apply heat to the tissue section 104 to attach it to the slide 106. The heat is preferably localized or mostly restricted to the top 108. For example, sufficient heat to cause attachment is supplied, in some embodiments, by heating the top 108 to a temperature of about 40°C to about 60°C. In some embodiments, a heat sink may be positioned in contact with the top of the slide to localize the applied heat to the top. In some embodiments, this method prevents the applied heat from diffusing below the slide, thus preventing more of the tissue section from adhering to the slide before smoothing.

[0036] Figure 2 shows another exemplary embodiment of the method and apparatus, similar to the embodiment in Figure 1, but with the heating element 210 located on the opposite side of the slide 206 from the tissue section 204. The tissue section 204 contains tissue 210 in an embedding medium 212 such as paraffin. The tissue collector 202 does not contain a heating element, but it is conceivable that the apparatus may have heaters / heating elements in the tissue collector 202 and heating element 210 on both sides of the slide. In Figure 2, the tissue collector 202 holds the tissue section 204, transports it from the microtome, and stretches the tissue section 204 toward the slide 206 until the tissue section 204 and the slide 206 are in physical contact. The heating element 210 applies heat to the slide 206, and the heat is transferred to the tissue section 204. As described with respect to Figure 1, the heat should be sufficient to cause attachment, either by itself or in combination with the physical force applied by the tissue collector 202, without damaging the slide.

[0037] Figure 3 is a perspective view of an exemplary embodiment of a device for automatically slicing tissue sections and transferring them to slides. The device comprises a rotary microtome 302, which can be an existing design or a new design adapted for use with the devices and methods of the present disclosure. For example, the rotary microtome 302 may comprise a base 304 and a microtome housing 306 provided on the base 304. A knife holder 308 is located on the base 304. A sample holder 310 can be moved up and down by a handwheel 312 provided on the microtome housing 306. The knife holder 308 and the sample holder 310 are positioned opposite each other. The knife holder 308 supports a front plate 316 (also called a knife pressure plate) that applies pressure to a knife 314. The knife holder 308 is supported by various clamps, supports and rails. The sample holder 310 can be adjusted to select the thickness of the slices to be taken from the tissue sample. The sample holder 310 also allows for adjustment of the angle at which the knife 314 contacts the tissue sample.

[0038] The apparatus operates as follows: The tissue sample is securely clamped or inserted into the sample holder 310 and moved around the knife 314 to lock it in place. The knife holder 308 is moved to the desired position for slicing tissue sections of the desired thickness and locked or clamped in place. The operator then rotates the handwheel 312 to lower the sample holder 310 so that the tissue sample engages with the knife 314. In Figure 3, the tissue section 341 has been sliced ​​by the microtome and is awaiting collection from the front plate 316.

[0039] The apparatus includes a tissue collector 332 that automatically collects tissue sections from the front plate 316. In Figure 3, the tissue sections 330 have been collected. The apparatus may include an automated tissue collector that moves relative to the knife so that the tissue collector can contact the sliced ​​tissue sections. For example, the tissue collector may be adapted so that (1) the collection end of the tissue collector moves to a position where it contacts or is sufficiently close to the tissue sections so that the tissue sections are collected by the tissue collector, and (2) the tissue sections on the collection end move to a position where they contact the slide held by the slide holder.

[0040] In some embodiments, the tissue collector is configured to collect tissue sections by suction, adhesion, friction, or other forces, rather than employing an adhesive material. For example, the tissue collector 332 may have one or more openings that provide a reduction in pressure (resulting in suction). The openings are fluidly connected to a pneumatic source. The tissue collector 332 may include an extendable portion, such as an arm 336, that contacts or approaches the tissue section 330. When the tissue collector 332 or a part thereof (such as an arm) is in contact with or close enough to the tissue section 330, the tissue section 330 is held in the tissue collector 332 by the reduction in pressure at the openings. "Reduced pressure" refers to a sub-ambient pressure that is lower than the ambient pressure, while "positive pressure" refers to a pressure that is higher than the ambient pressure, for example, greater than atmospheric pressure. The pressure gradient between the positive pressure and the ambient pressure propels the tissue section from the positive pressure region towards the reduced pressure region. "Suction" refers to the flow of gas into a partial vacuum or reduced pressure region. Due to the pressure gradient between this region and the ambient pressure, matter moves towards the reduced pressure region. In certain embodiments, sub-atmospheric pressure is a reduced pressure.

[0041] In some embodiments, the tissue collector employs an adhesive material instead of, or in addition to, reduced pressure. The adhesive material can be provided in any form, such as an adhesive strip. The term “strip,” as used herein, encompasses tapes, bands, patches, and other relatively flat forms. An adhesive preferred for use in tissue section collection is one that provides sufficient adhesion against movement but does not damage the tissue section when removed. Examples of such adhesives include low-tack pressure-sensitive adhesives, e.g., acrylonitrile copolymers (e.g., butadiene acrylonitrile polymer (BACN polymer), butadiene acrylonitrile isoprene polymer (BACNI polymer)); styrene copolymers (e.g., styrene / butadiene / styrene (SBS) polymer), styrene / isoprene / styrene (SIS polymer), and styrene / ethylene / butylene / styrene (SEBS polymer); and acrylate copolymers. Formulations and mixtures of polymer materials may be used as desired. The pressure-sensitive adhesive may also contain sufficient antioxidants, UV stabilizers, and crosslinking agents. Adhesive strips are available from various companies, including 3M Company in Maplewood, Minnesota, Polymer Science, Inc. in Monticello, Indiana, and Adhesives Research, Inc. in Glenrock, Pennsylvania.

[0042] The adhesive strip can be positioned on the tissue collector or on its stretchable portion by depressurization at the opening, by adhesive, by both, and / or by other means (i.e., by depressurization at the opening, by adhesive, by both, by other means, or all of them). For example, in Figure 3, the tissue collector 332 has an adhesive strip 338 positioned on the collection end. The tissue collector 332 includes a spool 337 on which the adhesive strip 338 can be unwound and rewound after being used to collect tissue sections 330. More specifically, a new adhesive strip is on a first reel 337 and is configured to extend across the collection end of the tissue collector 332. After a portion of the adhesive strip 338 has been used to collect tissue sections, the adhesive strip advances between reels, and the used portion of the adhesive strip is wound onto a second reel (not visible in Figure 3). The adhesive strip 338 comes into contact with the tissue section 330 and adheres to it. In some embodiments, the adhesive material contacts only a portion of the tissue section, preferably only the embedding medium and not the tissue. In other words, a first portion of the adhesive material is attached to the tissue section, while a second portion of the adhesive material is on the tissue collector but is not attached to or in contact with the tissue section.

[0043] The tissue collector 332 is designed to move so that it can collect tissue sections from a microtome or other location and transfer them to a slide. For example, the device may include an automated tissue collector mounted on a rotating support, the position of which changes as the rotating support rotates. In the embodiment shown in Figure 3, the tissue collector 332 is mounted on the support 342 by a shaft 340 so as to rotate. The support 342 can be operated manually or automatically by a controller, etc., and can rotate the shaft 340, which can accommodate the wiring of the tissue collector 332. The support 342 can move the tissue collector 332 to a first position, a second position, and beyond, where various operations are performed. As described above, the tissue collector 332 can be positioned to collect tissue sections 330 from a microtome. The tissue collector can then be rotated or otherwise moved to a position where it can transfer the tissue sections to a slide 344.

[0044] After acquiring a tissue section, the arm 336 moves so that the tissue section is positioned near the slide 344. The arm can be adapted to perform rotational and / or linear movements (i.e., rotational or linear movements or both). The arm or a portion thereof is extended again so that the tissue section can be attached to the slide by bringing the tissue section 330 and optionally the adhesive material into contact with the slide.

[0045] In Figure 3, the tissue collector 332 has rotated and the arm has extended, and the tissue section 330 and adhesive material 338 are moving toward contact with the slide 344 held by the slide holder 346. The tissue section 330 contacts the surface of the slide 344, and the adhesive strip 338 can be attached to the slide 344, in particular to the portion of the adhesive strip that is not attached to the tissue section. Heat and / or physical force (i.e., heat or physical force or both) can be applied to the tissue section 330 or a portion thereof, for example, at the contact point between the tissue section and the slide, for a sufficient amount and time to facilitate or increase the attachment of the tissue section 330 to the slide 344. The arm 336 retracts, and the tissue section 330 remains on the slide 344. In some embodiments, the adhesive strip 338 can also be left on the slide 344 by cutting or releasing both sides of the adhesive strip 338, or by simply stopping the depressurization that holds the adhesive material 338 in the tissue collector 332, thereby releasing the adhesive strip 338 from the tissue collector 332. In some embodiments, particularly when the adhesive strip 338 is provided on a reel, the adhesive material 338 is removed from the slide (manually or automatically as the arm 336 retracts), and the tissue section 330 remains attached to the slide 344 without adhesive.

[0046] After the tissue section has been transferred to the slide, the apparatus can be prepared for the next use. The tissue collector 332 can be rotated to the microtome 302 to collect the next tissue section 341, or otherwise moved. An adhesive dispenser can supply clean adhesive to the collection end. In embodiments where the adhesive is provided as a continuous strip, the used adhesive strip is removed from the collection end of the tissue collector, and a clean adhesive strip is moved onto the collection end. For example, if the adhesive strip is provided in a reel-to-reel configuration, for example, if the adhesive strip is provided on or wound onto a first reel, the adhesive strip is extended from the first reel across the collection end to the second reel, or rewound. The adhesive strip can be advanced across the collection end (partially unwound from the first reel and partially wound onto the second reel) after the first section of the adhesive strip has been used to transfer the tissue section onto the slide, thereby positioning the second section of the adhesive strip over the collection end. The tissue collector can be rotated or otherwise moved before, during, or after slicing the next tissue section. The tissue collector, slide holder, and slide can optionally be oriented along axes other than those shown in Figure 3 when the tissue section is mounted on the slide.

[0047] As described above, the tissue collector, or a part thereof, such as an arm, can be adapted to move to a position that contacts and / or collects a tissue section (i.e., contacts the tissue section, or collects it, or both). The outer surface of the tissue collector or arm can be flat, cylindrical, octagonal, or another shape, and can be adapted to perform additional movements. For example, the arm of the tissue collector can be adapted to extend and retract along a trajectory in the body of the tissue collector 332, and the arm 336 can be extended toward the tissue section or slide, come into contact with the tissue section or slide, and retract, thereby moving the collecting end of the tissue collector to contact the tissue section or close enough so that the tissue is collected by the tissue collector. In some embodiments, the arm can be rotatable, and this end can be rotated between a position that contacts and / or collects a tissue section (i.e., contacts the tissue section, or collects it, or both) and a position that places the tissue section on the slide. Optionally, the arm or tissue collector is configured to rotate to other positions where other functions or steps are performed.

[0048] In some embodiments of the apparatus, the slide is held by a slide holder 346, which may also be automated. The slide holder 346 may rotate along a slide holder base 350 or otherwise move, or may be mounted on the slide holder base 350 to provide such movement. For example, the slide holder base 350 may move along a track 352 toward the vessel 360. The slide holder 346 or slide holder base 350 may be adapted to move toward a position that contacts the vessel 360. In some embodiments, the slide holder 346 or slide holder base 350 is capable of rotation, which can change the angle of the slide 344. Optionally, the slide holder 346 or slide holder base 350 may be configured to rotate toward other positions where other functions or steps are performed. The apparatus in Figure 3 further comprises a slide holder base 350 and a controller 370 that signals to the tissue collector 332.

[0049] The slide holder can transport the slide 344 to the slide tank 360, which may contain hot water, or may be filled with cold water or room temperature water (or other liquid) first, and then with hot water (or other liquid). In some embodiments, the apparatus comprises two or more slide tanks, which may be at the same temperature or different temperatures, or contain the same or different liquids. For example, two separate water tanks may be provided, one at room temperature and the other at a high temperature. Other slide tank configurations may be used. The slide holder 346 can be positioned near one of the tissue collector locations and can be moved to immerse the held slide in the slide tank. Contacting the tissue section on the slide with hot water flattens the tissue section on the slide. The sequence of contacting the tissue section on the slide with cold water followed by hot water promotes flattening of the tissue section on the slide without bunching or bubbles. The adhesive strip can be removed (if not already removed) or left on the slide unless any part of the adhesive material interferes with the tissue analysis. Tissue sections flattened on the slide are generally ready for further processing (deparaffinization, stringency washing, coverslip mounting, staining, enzymatic treatment, etc.).

[0050] Figure 4 shows an exemplary embodiment of a tissue collector configured to have an adhesive strip at its collection end. The tissue collector 402 is capable of rotational and linear motion. The collection end 403 of the tissue collector 402 can be of any desired size or shape, for example, substantially flat with optionally rounded edges. The device also includes an adhesive material dispenser near the collection end 403 of the tissue collector 402, so that the collection end 403 of the tissue collector has an adhesive strip 405 on it. More specifically, the adhesive strip 405 extends across the collection end 403 from a first reel 412 to a second reel 416 across a wheel 414. The wheel 414 can be moved to tighten or loosen the adhesive strip 405 between the reel and the collection end. In some embodiments, the wheel 414 is a tachometer that provides a measure of how far the adhesive strip has advanced. A first reel 412 is wound with a clean, unused adhesive strip 405, and a second reel 416 is wound with a used adhesive strip. After the portion of the adhesive strip covering the collection end has been used to collect tissue or debris, the adhesive strip can be advanced by the reel, and the used adhesive strip is wound onto the second reel 416. Tension can be applied to the adhesive strip using a variety of additional elements different from those shown in Figure 4. In some embodiments, the apparatus includes an actuator that clamps an unused adhesive strip to the surface of the collection end 403 (such as the top or side facing the slide) if the wheel 414 has not wound more adhesive strip onto the spool over the collection end 403.

[0051] The tissue collector 402 is capable of linear motion toward the slide 406, and when a tissue section is collected at the collection end 403, the tissue collector can bring the tissue section into contact with the slide 406 held by the slide holder 418. On the opposite side of the slide 406, there is a heating element 410 which is capable of linear motion toward the slide 406. The heating element 410 can be maintained at a desired temperature or can be configured to heat up quickly when desired for use. The heating element 410 contacts the surface of the slide 406 opposite to the side that contacts the tissue section. The heating element 410 can be configured to contact only a portion of the slide. Applying heat from the opposite side reduces the risk of damaging the tissue section.

[0052] The heating element 410 may remain stationary while the slide holder 406 and the tissue collector 402 move to bring them both into contact. The heating element 410 may be omitted, and / or the collection end 403 may be equipped with a heating element (i.e., the heating element 410 may be omitted, or the collection end 403 may be equipped with a heating element, or both), and heat may be applied to the tissue section from the collection end 403. The surfaces of the tissue collector 402 and the heating element 410 facing the tissue section may be configured to rotate, for example, by having a pivot axis 422 in the tissue collector 402 and a pivot axis 420 in the heating element 410.

[0053] By using the apparatus and method described herein, tissue sections can be automatically transferred from a microtome to a slide without manual handling of the tissue sections. The tissue sections can be transferred without manual contact with them, for example, without the user grasping the tissue sections by hand or with a handheld instrument. More specifically, freshly sliced ​​tissue sections can be removed from the microtome and / or placed on a slide without manual contact (i.e., freshly sliced ​​tissue sections can be removed from the microtome, or placed on a slide, or both, without manual contact). Furthermore, the apparatus and method offer good two-dimensional stability, enabling slicing and transfer of tissue sections with bunching or curling, and avoiding tissue section waste.

[0054] [Example Embodiments] Exemplary embodiments provided by the subject matter disclosed herein include, but are not limited to, the following:

[0055] Embodiment 1. A method for transferring a tissue section onto a slide, comprising: collecting the tissue section using a tissue collector and / or adhesive strip; moving the tissue collector and / or adhesive strip to bring at least a portion of the tissue section into contact with the slide; and applying physical force, heat, or both to the tissue section at the point of contact between the tissue section and the slide.

[0056] Embodiment 2. The method according to Embodiment 1, further comprising heating the contact area of ​​the tissue section to a temperature of approximately 40°C to approximately 90°C.

[0057] Embodiment 3. The method according to Embodiment 1 or 2, wherein the tissue section comprises an embedding medium, and the heat is sufficient to at least partially melt the embedding medium.

[0058] Embodiment 4. The method according to any one of Embodiments 1 to 3, comprising applying a physical force of approximately 275 kPA or less to the contact area of ​​the tissue section.

[0059] Embodiment 5. The method according to any one of Embodiments 1 to 4, wherein the contact area occupies 25% or less of the tissue section.

[0060] Embodiment 6. The method according to any one of Embodiments 1 to 5, further comprising positioning the adhesive strip on the collection end of the tissue collector.

[0061] Embodiment 7. The method according to Embodiment 6, comprising collecting the tissue sections by adhering them to the adhesive strip.

[0062] Embodiment 8. The method according to Embodiment 6, wherein the adhesive strip is provided on a first reel, and the adhesive strip extends from the first reel across the collection end to a second reel.

[0063] Embodiment 9. The method according to Embodiment 8, wherein, after the first section of the adhesive strip has been used to transfer a tissue section to a slide, the adhesive strip is advanced across the collection end (a portion is unwound from the first reel and a portion is wound onto the second reel), thereby positioning the second section of the adhesive strip over the collection end.

[0064] Embodiment 10. The method according to any one of Embodiments 1 to 9, wherein the adhesive strip comprises a low-tack pressure-sensitive adhesive.

[0065] Embodiment 11. Apparatus for transferring tissue sections to a slide, comprising: a tissue collector for collecting the tissue sections; a slide holder for holding the slide, wherein the tissue collector and / or the slide holder is configured to move such that the tissue sections collected by the tissue collector and the slide held by the slide holder come into contact with each other at a contact point; and a heating element configured to be positioned at the contact point or to move to the contact point.

[0066] Embodiment 12. The apparatus according to Embodiment 11, wherein the heating element is housed within the tissue collector.

[0067] Embodiment 13. The apparatus according to Embodiment 11, wherein the heating element is housed in the slide holder.

[0068] Embodiment 14. The apparatus according to any one of Embodiments 11 to 13, wherein the heating element is located outside the tissue section and the slide holder, the heating element is positioned on a first surface of the slide held by the slide holder or is movable to a position on the first surface, and the contact point is a second opposing surface of the slide.

[0069] Embodiment 15. The apparatus according to any one of Embodiments 11 to 14, further comprising a dispenser for adhesive material.

[0070] Embodiment 16. Apparatus for transferring tissue sections to a slide, comprising: a tissue collector for collecting the tissue sections; a slide holder for holding the slide, wherein the tissue collector and / or the slide holder are configured to move such that the tissue sections collected by the tissue collector and the slide held by the slide holder come into contact with each other at contact points; and an adhesive strip dispenser capable of supplying an adhesive strip at the collection end of the tissue collector.

[0071] Embodiment 17. The apparatus according to Embodiment 16, wherein the adhesive strip dispenser comprises at least two reels on both sides of the collection end, the adhesive strip extends from a first reel across the collection end to a second reel, the first reel is capable of unwinding new adhesive strips, and the second reel is capable of winding used adhesive strips.

[0072] Embodiment 18. The apparatus according to Embodiment 17, wherein the collection end of the tissue collector is extendable.

[0073] Embodiment 19. The apparatus according to Embodiment 17 or 18, wherein the tissue collector is configured to transfer the tissue section by applying physical force and / or heat to or near the slide holder.

[0074] Embodiment 20. The apparatus according to any one of Embodiments 16 to 19, wherein the tissue collector or the slide holder is equipped with a heating element.

[0075] The above description of exemplary or preferred embodiments should be interpreted as illustrative and not as limiting the invention as defined by the embodiments. For ease of understanding, numerous variations and combinations of the features described above can be used without departing from the invention as described in the embodiments. Such variations will not be considered departures from the scope of the invention, and all such variations are intended to be included within the scope of the following embodiments. All references cited herein constitute, by reference, part in whole of this specification.

Claims

1. The tissue sections are collected by holding them in a tissue collector, The tissue collector is moved to bring a portion of the tissue section that does not contain tissue into contact with the slide, Applying physical force, heat, or both to the tissue section at the point of contact between the tissue section and the slide. A method for transferring tissue sections onto a slide, including [details omitted].

2. The method according to claim 1, comprising heating the contact area of ​​the tissue section to a temperature of 40°C to 90°C.

3. The method according to claim 1 or 2, wherein the tissue section comprises an embedding medium, and the heat is sufficient to at least partially melt the embedding medium.

4. The method according to any one of claims 1 to 3, comprising applying a physical force of 275 kPa or less to the contact area of ​​the tissue section.

5. The method according to any one of claims 1 to 4, wherein the contact area occupies 25% or less of the tissue section.

6. The method according to any one of claims 1 to 5, further comprising positioning the adhesive strip on the collection end of the tissue collector.

7. The method according to claim 6, comprising collecting the tissue section by adhering it to the adhesive strip.

8. The method according to claim 6, wherein the adhesive strip is provided on a first reel, and the adhesive strip extends from the first reel across the collection end to a second reel.

9. The method according to claim 8, comprising advancing the adhesive strip across the collection end after the first section of the adhesive strip has been used to transfer a tissue section to a slide, thereby positioning the second section of the adhesive strip on the collection end.

10. The method according to any one of claims 6 to 9, wherein the adhesive strip comprises a low-tack pressure-sensitive adhesive.

11. A tissue collector configured to collect tissue sections by holding the tissue sections, A slide holder designed to hold the slide and Equipped with, At least one of the tissue collector and the slide holder is configured to move such that a portion of the tissue section collected by the tissue collector that does not contain tissue and the slide held by the slide holder come into contact with each other. A device for transferring a tissue section to a slide, further comprising a heating element configured to be positioned at the contact point between the portion of the tissue section and the slide, or to move to the contact point.

12. The apparatus according to claim 11, wherein the heating element is housed in the tissue collector.

13. The apparatus according to claim 11, wherein the heating element is housed in the slide holder.

14. The apparatus according to any one of claims 11 to 13, wherein the heating element is located outside the tissue section and the slide holder, the heating element is positioned on a first surface of the slide held by the slide holder or is movable to a position on the first surface, and the contact point is a second opposing surface of the slide.

15. The apparatus according to any one of claims 11 to 14, further comprising a dispenser for adhesive material.

16. A tissue collector that collects tissue sections by holding the tissue sections at its collection end, A slide holder designed to hold the slide, Equipped with, At least one of the tissue collector and the slide holder is configured to move such that a portion of the tissue section collected by the tissue collector that does not contain tissue and the slide held by the slide holder come into contact with each other. At least one of the tissue collector and the slide holder is configured to apply physical force, heat, or both to the contact point between the portion of the tissue section and the slide. A device for transferring tissue sections to a slide, further comprising an adhesive strip dispenser capable of supplying an adhesive strip at the collection end of the tissue collector.

17. The apparatus according to claim 16, wherein the adhesive strip dispenser comprises at least two reels on either side of the collection end, the adhesive strip extends from a first reel across the collection end to a second reel, the first reel is capable of unwinding new adhesive strips, and the second reel is capable of winding used adhesive strips.

18. The apparatus according to claim 17, wherein the collection end of the tissue collector is extendable.

19. The apparatus according to claim 17 or 18, wherein the tissue collector is configured to transfer the tissue section by applying physical force and / or heat to or near the slide holder.

20. The apparatus according to any one of claims 16 to 19, wherein the tissue collector or the slide holder is equipped with a heating element.

Images