Automated tissue section transport system with high throughput

Abstract

The present disclosure relates to automated systems and methods for sectioning tissue from biological tissue blocks, and more particularly to systems and methods for providing higher throughput in such automated devices. The present disclosure can implement systems and methods for increasing throughput in automated tissue sectioning devices to reduce tissue sample preparation time for multiple tissue blocks. The present disclosure further provides systems and methods for hydrating tissue samples.

Description

[Technical Field]

[0001] (CROSS-REFERENCE TO RELATED APPLICATIONS) This application claims the benefit of and priority to U.S. Provisional Application No. 63 / 075,441, filed September 8, 2020, U.S. Provisional Application No. 63 / 077,433, filed September 11, 2020, U.S. Utility Application No. 17 / 469,351, filed September 8, 2021, and U.S. Utility Application No. 17 / 469,364, filed September 8, 2021, all of which are incorporated herein by reference in their entireties.

[0002] The present disclosure relates to automated systems and methods for sectioning tissue from biological tissue blocks, and more particularly to systems and methods that provide higher throughput in such automated devices. [Background technology]

[0003] Traditional microtomy, the production of micron-thin tissue sections for microscopic viewing, is a delicate and time-consuming manual task. Recent advances in digital imaging of tissue sample sections have made it desirable to slice specimen blocks very quickly. As an example, when tissue is sectioned as part of a clinical procedure, time is a critical variable in improving patient treatment. For example, every minute that can be saved during sectioning of tissue for intraoperative use in anatomical pathology, such as examining the margins of a lung cancer to determine whether sufficient tissue has been removed, is clinically beneficial. To rapidly produce large numbers of sample sections, it is desirable to automate the process of cutting tissue sections from supporting tissue blocks with a microtome blade and facilitating the transfer of the cut tissue sections to slides.

[0004] Any time that can be saved during sectioning of tissue for intraoperative applications in anatomical pathology can be important. It would be advantageous to provide an automated system that could increase tissue sectioning and save time. Summary of the Invention [Means for solving the problem]

[0005] A need exists for improved systems and methods for tissue sample preparation. The present disclosure is directed to a solution to address this need, in addition to possessing other desirable properties.

[0006] In some aspects, the present disclosure provides an automated system for preparing tissue samples, comprising one or more microtomes, a hydration system, and a processor, the processor being programmed to: initiate sectioning of a first tissue block with the one or more microtomes, the first tissue block comprising a first tissue sample embedded within an embedding material; hydrating the first tissue block with the hydration system for a first predetermined time; initiate sectioning of a second tissue block with the one or more microtomes while the first tissue block is hydrated, the second tissue block comprising a second tissue sample embedded within an embedding material, hydrating the second tissue block with the hydration system for a second predetermined time; and initiate the one or more microtomes to begin sectioning of the first tissue block while the second tissue block is hydrated.

[0007] In some embodiments, the one or more microtomes comprise a first microtome and a second microtome. In some embodiments, the method can further include the automated system of claim 1, wherein the processor is programmed to: initiate sectioning of a first tissue block with the first microtome, hydrate the first tissue block with the hydration system, initiate sectioning of a second tissue block with the first microtome while the first tissue block is in the hydration system, hydrate the second tissue block with the hydration system, and initiate sectioning of the hydrated first tissue block with the second microtome while the second tissue block is hydrated. In some embodiments, the processor is further programmed to initiate sectioning of the first tissue with the first microtome, hydrating the first tissue block with the hydration system, initiating sectioning of the hydrated first tissue block with the first microtome, sectioning the second tissue block with the second microtome, hydrating the second tissue block with the hydration system, and initiating sectioning of the hydrated second tissue block with the second microtome, wherein at least one of the sectioning, hydration, and sectioning with the second microtome is performed in parallel with the sectioning, hydration, and sectioning with the first microtome.

[0008] In some embodiments, the system further includes one or more transport medium units configured to transfer one or more sections from the first and second tissue blocks to one or more slides. In some embodiments, the system further includes one or more tissue block handlers configured to transfer the first and second tissue blocks between the one or more microtomes and the hydration system. In some embodiments, the system further includes a processor programmed to communicate with the one or more tissue block handlers and control movement of the first and second tissue blocks between the one or more microtomes and the hydration system. In some embodiments, the system further includes a tissue block tray configured to receive the first and second tissue blocks. In some embodiments, the system further includes a cooling system. In some embodiments, the system further includes a hydration chamber configured to receive the first and second tissue blocks inside the hydration chamber. In some embodiments, the system can further include a hydration system positioned proximate to the one or more microtomes and configured to hydrate the first and second tissue blocks on the one or more microtomes. The processor can be further programmed in some embodiments to move the first and second tissue blocks through the hydration system for a first predetermined time and a second predetermined time, respectively.

[0009] In some aspects, the present disclosure also provides a method for batch processing tissue blocks. In some embodiments, the method involves initiating, by a processor, cross-sectioning of a first tissue block with one or more microtomes controlled by the processor, the first tissue block comprising a first tissue sample embedded in an embedding material, hydrating, by the processor, the first tissue block for a first predetermined period of time, initiating, by the processor, cross-sectioning of a second tissue block with the one or more microtomes while the first tissue block is hydrated, the second tissue block comprising a second tissue sample embedded in an embedding material, hydrating, by the processor, the second tissue block for a second predetermined period of time, and initiating, by the processor, the one or more microtomes to begin sectioning the first tissue block while the second tissue block is hydrated. In some embodiments, the method may further include transferring the first and second tissue blocks from the one or more microtomes to a hydration system by a tissue block handler in communication with the processor. In some embodiments, the one or more microtomes may comprise multiple microtomes. In some embodiments, the method may further include transferring one or more sections sectioned from the first and second tissue blocks from the one or more microtomes to one or more slides by one or more transport medium units in communication with the processor.

[0010] In some aspects, the present disclosure also provides a method for processing a tissue sample, comprising: sectioning, with a first microtome, a first tissue block comprising a first tissue sample embedded in an embedding material; hydrating the first tissue block with a hydration system for a predetermined period of time; sectioning, with the first microtome, a second tissue block comprising the second tissue sample embedded in an embedding material while the first tissue block is hydrated; hydrating the second tissue block with the hydration system; and sectioning, with the second microtome, a sample from the hydrated first tissue block while the second tissue block is hydrated. In some embodiments, the method further comprises transferring one or more sections sectioned from the first tissue block from the second microtome to one or more slides.

[0011] In some aspects, the present disclosure provides a method for processing a tissue sample, comprising: sectioning, with a first microtome, a first tissue block comprising a first tissue sample embedded in an embedding material; hydrating the first tissue block with a hydration system for a first predetermined period of time; sectioning the hydrated first tissue block with the first microtome; sectioning, with a second microtome, a second tissue block comprising a second tissue sample embedded in an embedding material; hydrating the second tissue block with a hydration system for a second predetermined period of time; and sectioning the hydrated second tissue block with the second microtome, wherein at least one of the sectioning, hydration, and sectioning with the second microtome is performed in parallel with the sectioning, hydration, and sectioning with the first microtome. In some embodiments, the method may further include transferring one or more sections sectioned from the first tissue block from the first microtome and the second microtome to one or more slides.

[0012] In some aspects, the present disclosure provides a histology system comprising: a microtome configured to expose a face of a tissue block comprising a tissue sample embedded in an embedding material and remove one or more tissue sections from the sample; a hydration system configured to hydrate the tissue block by depositing a hydrating liquid on the face of the tissue block; and a transfer system configured to transfer the one or more tissue sections from the microtome to one or more slides. In some embodiments, the hydration system is configured to generate droplet condensation of the hydrating liquid and deposit the condensation on the face of the tissue block. In some embodiments, the hydration system comprises an ultrasonic humidifier. In some embodiments, the hydration system comprises a sprayer assembly configured to spray droplets of the hydrating liquid onto the face of the tissue block. In some embodiments, the hydration system comprises a hydration chamber configured to receive one or more tissue blocks and a humidifier in communication with the hydration chamber to provide humid air to the hydration chamber. In some embodiments, the hydrating liquid is deposited by spraying droplets of the hydrating liquid onto the surface of the tissue sample block. In some embodiments, the hydrating liquid comprises a surfactant. In some embodiments, the surfactant is an oil-soluble surfactant. In some embodiments, the surfactant is a superdiffusing surfactant. In some embodiments, the method further comprises a cooling system configured to cool the tissue block. In some embodiments, the hydration system is positioned proximate to the microtome and configured to hydrate the tissue block in the microtome.

[0013] In some aspects, the present disclosure also provides a method for hydrating a tissue block, comprising: exposing a face of a tissue block comprising a tissue sample embedded in an embedding material; depositing a hydrating liquid on the face of the tissue block; allowing the hydrating liquid to hydrate the tissue sample; and sectioning one or more tissue sections from the hydrated tissue sample. In some embodiments, the hydrating liquid is deposited by causing condensation of the hydrating liquid on the face of the tissue block. In some embodiments, the hydrating liquid is deposited by spraying droplets of the hydrating liquid onto the face of the tissue sample block. In some embodiments, the hydrating liquid comprises a surfactant. In some embodiments, the surfactant is an oil-soluble surfactant. In some embodiments, the surfactant is a super-diffusing surfactant.

[0014] In some embodiments, the method further includes positioning a tissue block in a microtome chuck adjacent to a microtome configured to section one or more sections from the hydrated tissue sample, and depositing a hydration liquid on a surface of the tissue sample positioned on the microtome chuck. In some embodiments, the method further includes positioning the tissue block in a hydration chamber and providing air with vapor of the hydration liquid to the hydration chamber such that droplets of the hydration liquid condense on the surface of the tissue block positioned in the hydration chamber. In some embodiments, the method further includes cooling the tissue block to induce droplets of the hydration liquid to condense on the surface of the tissue block.

[0015] These and other embodiments of the present disclosure are described in more detail below. The present specification also provides, for example, the following: (Item 1) 1. An automated system for preparing a tissue sample, the system comprising: one or more microtomes; A hydration system; A processor, the processor comprising: commencing sectioning of a first tissue block with one or more microtomes, the first tissue sample being embedded within an embedding material; hydrating the first tissue block with the hydration system for a first predetermined time period; commencing sectioning of a second tissue block with the one or more microtomes while the first tissue block is hydrated, the second tissue block comprising a second tissue sample embedded within an embedding material; hydrating the second tissue block with the hydration system for a second predetermined time period; starting the one or more microtomes to begin sectioning the first tissue block while the second tissue block is hydrated; a processor programmed to A system comprising: (Item 2) Item 10. The automated system of item 1, wherein the one or more microtomes comprise a first microtome and a second microtome. (Item 3) The processor: commencing sectioning of the first tissue block with a first microtome; hydrating the first tissue block with the hydration system; commencing sectioning of the second tissue block with the first microtome while the first tissue block is within the hydration system; hydrating the second tissue block with the hydration system; commencing sectioning of the hydrated first tissue block with a second microtome while the second tissue block is hydrated; Item 1. The automated system according to item 1, programmed to: (Item 4) The processor: commencing sectioning of the first tissue block with a first microtome; hydrating the first tissue block with the hydration system; commencing sectioning of the hydrated first tissue block with the first microtome; sectioning the second tissue block with a second microtome; hydrating the second tissue block with the hydration system; commencing sectioning of the hydrated second tissue block with the second microtome; It is programmed to Item 10. The automated system of item 1, wherein at least one of the sectioning, hydration, and sectioning by the second microtome is performed in parallel with the sectioning, hydration, and sectioning by the first microtome. (Item 5) 5. The automated system of any one of items 1-4, further comprising one or more transfer medium units configured to transfer one or more sections from the first tissue block and the second tissue block to one or more slides. (Item 6) 5. The automated system of any one of items 1-4, further comprising one or more tissue block handlers configured to transfer the first tissue block and the second tissue block between the one or more microtomes and the hydration system. (Item 7) 7. The automated system of claim 6, wherein the processor is programmed to communicate with the one or more tissue block handlers and control movement of the first tissue block and the second tissue block between the one or more microtomes and the hydration system. (Item 8) 5. The automated system of any one of items 1-4, further comprising a tissue block tray configured to receive the first tissue block and the second tissue block. (Item 9) 5. The automated system of any one of items 1-4, further comprising a cooling system. (Item 10) Item 10. The automated system of item 1, wherein the hydration system comprises a hydration chamber configured to receive the first tissue block and the second tissue block inside the hydration chamber. (Item 11) Item 11. The automated system of item 10, wherein the processor is further programmed to move the first tissue block and the second tissue block to the hydration system for the first predetermined time and the second predetermined time, respectively. (Item 12) 5. The automated system of any one of items 1-4, wherein the hydration system is positioned proximate to the one or more microtomes and configured to hydrate the first tissue block and the second tissue block in the one or more microtomes. (Item 13) 1. A method for batch processing tissue blocks, the method comprising: initiating, by a processor, sectioning of a first tissue block comprising a first tissue sample embedded within an embedding material by one or more microtomes controlled by said processor; hydrating, by the processor, the first tissue block for a first predetermined period of time; initiating, by the processor, sectioning with the one or more microtomes a second tissue block while the first tissue block is hydrated, the second tissue block comprising a second tissue sample embedded within an embedding material; hydrating the second tissue block by the processor for a second predetermined period of time; starting, by the processor, the one or more microtomes to begin sectioning the first tissue block while the second tissue block is hydrated; A method comprising: (Item 14) 14. The method of claim 13, further comprising transferring, by a tissue block handler in communication with the processor, the first tissue block and the second tissue block from the one or more microtomes to a hydration system for hydration. (Item 15) Item 14. The method of item 13, wherein the one or more microtomes comprise a plurality of microtomes. (Item 16) 14. The method of claim 13, further comprising transferring, by one or more transport medium units in communication with the processor, one or more sections sectioned from the first tissue block and the second tissue block from the one or more microtomes to one or more slides. (Item 17) 1. A method for processing a tissue sample, the method comprising: sectioning, with a first microtome, a first tissue block comprising a first tissue sample embedded within an embedding material; hydrating the first tissue block with a hydration system for a predetermined period of time; sectioning a second tissue block with the first microtome while the first tissue block is hydrated, the second tissue block comprising a second tissue sample embedded in an embedding material; hydrating the second tissue block with the hydration system; sectioning a sample from the hydrated first tissue block with a second microtome while the second tissue block is hydrated; A method comprising: (Item 18) 18. The method of claim 17, further comprising transferring one or more sections sectioned from the first tissue block from the second microtome to one or more slides. (Item 19) 1. A method for processing a tissue sample, the method comprising: sectioning, with a first microtome, a first tissue block comprising a first tissue sample embedded within an embedding material; hydrating the first tissue block with a hydration system for a first predetermined period of time; sectioning the hydrated first tissue block with the first microtome; sectioning, with a second microtome, a second tissue block comprising a second tissue sample embedded within an embedding material; hydrating the second tissue block with the hydration system for a second predetermined period of time; sectioning the hydrated second tissue block with the second microtome; Including, A method wherein at least one of the sectioning, hydrating, and sectioning with the second microtome is performed in parallel with the sectioning, hydrating, and sectioning with the first microtome. (Item 20) 20. The method of claim 19, further comprising transferring one or more sections sectioned from the first tissue block from the first microtome and the second microtome to one or more slides. (Item 21) 1. A system for histology, comprising: a microtome configured to expose a face of a tissue block comprising a tissue sample embedded within an embedding material and remove one or more tissue sections from said sample; a hydration system configured to hydrate the tissue block by depositing a hydrating liquid on a surface of the tissue block; a transfer system configured to transfer the one or more tissue sections from the microtome to one or more slides; A system comprising: (Item 22) 22. The system of claim 21, wherein the hydration system is configured to generate droplet condensations of the hydration liquid and deposit the droplet condensations on the surface of the tissue block. (Item 23) 22. The system of claim 21, wherein the hydration system comprises an ultrasonic humidifier. (Item 24) 22. The system of claim 21, wherein the hydration system comprises a sprayer assembly configured to spray droplets of the hydration liquid onto the surface of the tissue block. (Item 25) Item 22. The system of item 21, wherein the hydration system comprises a hydration chamber configured to receive one or more tissue blocks and a humidifier in communication with the hydration chamber to provide moist air to the hydration chamber. (Item 26) 22. The system of claim 21, wherein the hydrating liquid is deposited by spraying droplets of the hydrating liquid onto the surface of the tissue block. (Item 27) 22. The system of claim 21, wherein the hydrating liquid comprises a surfactant. (Item 28) 28. The system according to claim 27, wherein the surfactant is an oil-soluble surfactant. (Item 29) 28. The system of claim 27, wherein the surfactant is a super-diffusing surfactant. (Item 30) 30. The system of any one of items 21-29, further comprising a cooling system configured to cool the tissue block. (Item 31) 30. The system of any one of items 21-29, wherein the hydration system is positioned adjacent to the microtome and configured to hydrate the tissue block in the microtome. (Item 32) 1. A method for tissue block hydration, said method comprising: exposing a face of a tissue block comprising a tissue sample embedded within an embedding material; depositing a hydrating liquid on a surface of the tissue block; allowing the hydrating liquid to hydrate the tissue sample; and sectioning one or more tissue sections from the hydrated tissue sample. A method comprising: (Item 33) 33. The method of claim 32, wherein the hydrating liquid is deposited by causing condensation of the hydrating liquid on the surface of the tissue block. (Item 34) 33. The method of claim 32, wherein the hydrating liquid is deposited by spraying droplets of the hydrating liquid onto the surface of the tissue block. (Item 35) 33. The method of claim 32, wherein the hydrating liquid comprises a surfactant. (Item 36) Item 36. The method according to item 35, wherein the surfactant is an oil-soluble surfactant. (Item 37) Item 36. The method of item 35, wherein the surfactant is a super-diffusing surfactant. (Item 38) positioning the tissue block in a microtome chuck adjacent to a microtome, the microtome configured to section one or more sections from the hydrated tissue sample; depositing the hydrating liquid onto a surface of the tissue sample positioned on the microtome chuck; 38. The method according to any one of items 32-37, further comprising: (Item 39) Positioning the tissue block within a hydration chamber; providing air carrying vapor of the hydration liquid into the hydration chamber so that the hydration liquid condenses on the surface of the tissue block positioned within the hydration chamber; 38. The method according to any one of items 32-37, further comprising: (Item 40) 40. The method of claim 39, further comprising cooling the tissue block to induce droplets of the hydrating liquid to condense on the surface of the tissue block. [Brief explanation of the drawings]

[0016] The present disclosure is further described in the detailed description that follows with reference to several drawings in which like reference numerals represent similar parts throughout the several views of the drawings, and in which are set forth non-limiting examples of illustrative embodiments.

[0017] [Figure 1A] FIG. 1A is a top view illustration of a sample system layout according to some embodiments of the present disclosure.

[0018] [Figure 1B] 1B and 1C are isometric illustrations of a sample system layout according to some embodiments of the present disclosure. [Figure 1C] 1B and 1C are isometric illustrations of a sample system layout according to some embodiments of the present disclosure.

[0019] [Figure 2] FIG. 2 is a flow chart illustrating batch processing for increasing throughput in an automated sectioning system, according to some embodiments of the present disclosure.

[0020] [Figure 3] FIG. 3 is a chart comparing batch and continuous block processing, with batch processing shown to the left of the black line and continuous processing shown to the right of the black line, in accordance with some embodiments of the present disclosure.

[0021] [Figure 4A] 4A and 4B are flow charts illustrating the operation of multiple microtomes in tandem, according to some embodiments of the present disclosure. [Figure 4B] 4A and 4B are flow charts illustrating the operation of multiple microtomes in tandem, according to some embodiments of the present disclosure.

[0022] [Figure 5A] 5A and 5B are flow charts illustrating the operation of multiple microtomes in parallel, according to some embodiments of the present disclosure. [Figure 5B] 5A and 5B are flow charts illustrating the operation of multiple microtomes in parallel, according to some embodiments of the present disclosure.

[0023] [Figure 6] FIG. 6 is a graph illustrating block hydration capacity using a single microtome in one example of a multiple pass process, according to some embodiments of the present disclosure.

[0024] [Figure 7] 7A and 7B are graphs illustrating block hydration volumes using two microtomes in tandem (upper graph) and parallel (lower graph) operation, according to some embodiments of the present disclosure.

[0025] [Figure 8] FIG. 8 is a flow chart illustrating an exemplary hydration process of the present disclosure.

[0026] [Figure 9] 9-11 illustrate exemplary components for batch processing of tissue blocks according to the present disclosure. [Figure 10] 9-11 illustrate exemplary components for batch processing of tissue blocks according to the present disclosure. [Figure 11] 9-11 illustrate exemplary components for batch processing of tissue blocks according to the present disclosure.

[0027] [Figure 12] FIG. 12 illustrates exemplary components for hydrating individual tissue blocks according to the present disclosure.

[0028] [Figure 13] FIG. 13 is an exemplary high-level architecture for implementing a process according to the present disclosure.

[0029] While the above-identified drawings set forth embodiments of the presently disclosed subject matter, other embodiments are also contemplated, as noted in the discussion. The present disclosure presents illustrative embodiments by way of representation, not limitation. Numerous other modifications and embodiments can be devised by those skilled in the art which fall within the scope and spirit of the principles of the presently disclosed subject matter. DETAILED DESCRIPTION OF THE INVENTION

[0030] Detailed Description The present disclosure relates to systems and methods for processing tissue blocks containing biological samples of tissue. Processing can include automated systems designed to section the tissue block and cut tissue sections from the tissue block. The cut tissue sections can be transferred to a transfer / transport medium, such as tape, and then transferred from the transfer medium to slides for pathology or histology examination. The disclosed methods and systems can be employed in conjunction with manual and automated microtomy methods and systems.

[0031] The present disclosure can implement systems and methods for increasing throughput in automated tissue sectioning devices to reduce tissue sample preparation time for multiple tissue blocks. In some embodiments, the present disclosure implements automated systems and methods that maximize the number of tissue blocks processed by coordinating various processing steps in an efficient manner and minimizing delays in tissue sample preparation. The present disclosure provides several systems and methods that can improve the speed of automated devices, and these systems and methods can be used alone or in combination.

[0032] In some embodiments, the systems and methods of the present disclosure can implement automated equipment with more than one microtome, so that more than one path through the system can be utilized in tandem and / or parallel. It should be understood that providing more than one microtome for sectioning or sectioning a tissue block is one way in which multiple paths (redundancy) for the same task can be provided, and other redundancies can be incorporated into the system / process. In some embodiments, the systems and methods of the present disclosure can include parallel processing, i.e., moving blocks from a chuck to a block holder and preparing slides simultaneously. In some embodiments, tissue blocks can be processed in tandem.

[0033] In some embodiments, the present disclosure provides a method for batch processing sample blocks, the method including the steps of sectioning a block containing a biological sample with a microtome, moving a first tissue block into a hydration chamber for a predetermined period of time, sectioning the sample from the hydrated block with the microtome, and transferring the sample from the block to a transport medium for analysis.

[0034] In some embodiments, the present disclosure provides a tandem method for batch processing sample blocks, the method including: sectioning a first tissue block containing a first biological sample with a first microtome; moving the first tissue block into a hydration chamber for a predetermined period of time; sectioning a second tissue block containing a second biological sample with the first microtome while the first tissue block is in the hydration chamber; moving the second tissue block into the hydration chamber; sectioning a sample from the hydrated first tissue block with the second microtome while the second tissue block is in the hydration chamber; and transferring the sample from the first tissue block to a transport medium for analysis.

[0035] In some embodiments, the present disclosure provides a parallel method for batch processing sample blocks, the method including the steps of: sectioning a first tissue block containing a first biological sample with a first microtome; transferring the first tissue block into a hydration chamber for a predetermined period of time; sectioning the first sample from the hydrated first tissue block with the first microtome; transferring the first sample from the block to a transfer medium for analysis; sectioning a second tissue block containing a second biological sample with a second microtome; transferring the second tissue block into the hydration chamber for a predetermined period of time; sectioning a second sample from the hydrated second tissue block with the second microtome; and transferring the second sample from the second tissue block to a transfer medium for analysis, wherein at least one of the sectioning, hydration, and sectioning with the second microtome is performed in parallel with the sectioning, hydration, and sectioning with the first microtome.

[0036] The present disclosure further provides methods and systems for enhanced hydration of tissue blocks during microtomy. In some embodiments, the tissue blocks can be hydrated in a hydration chamber. In some embodiments, the tissue blocks can be hydrated directly in the microtome.

[0037] 1A-13, in which like parts are designated by like reference numerals throughout, illustrate an exemplary embodiment or embodiments of improved operations for processing tissue blocks containing biological samples according to the present disclosure. While the present disclosure will be described with reference to the exemplary embodiment or embodiments illustrated in the figures, it should be understood that many alternative forms may embody the present disclosure. Those skilled in the art will additionally understand different ways to modify parameters of the disclosed embodiments, such as the size, shape, or type of elements or materials, in a manner that remains consistent with the spirit and scope of the present disclosure.

[0038] In some embodiments, the present disclosure can be used in conjunction with tissue blocks containing biological samples such as tissue. The systems and methods of the present disclosure can be used to efficiently process and separate tissue blocks. Tissue samples are typically embedded in a preservative material, such as paraffin wax or a similar material. The embedding process can include any combination of processes for producing tissue blocks designed to be cut by the microtome 104. For example, the biological sample can be enclosed in a mold with a liquid substance, such as wax or epoxy, which can harden to produce a block of the desired shape. Once the tissue blocks are created, they can be inserted into the automated system 100 for cutting into samples that can be placed on slides for observation.

[0039] In particular, as discussed in detail below, the automated system 100 is designed to receive one or more tissue blocks, each comprising a tissue sample embedded in an embedding or preservation material. The tissue blocks are delivered to one or more microtomes. The one or more tissue blocks are then "sectioned" by removing the layer of preservation material in which the tissue is embedded, exposing a large cross-section of the tissue sample, e.g., the front surface of the tissue sample. Such exposed surface of the tissue sample in the tissue block is referred to as the block face. Once the tissue block is sectioned, the tissue block can be hydrated and cooled prior to sectioning the tissue block (cutting tissue sections that can be mounted on slides for observation). The tissue block can be hydrated and cooled in a separate hydration chamber or can be hydrated and cooled in place in one or more microtomes. The sections of the tissue sample are transferred from the one or more microtomes to slides for further processing.

[0040] 1A, 1B, and 1C, in some embodiments, an automated pathology system 100 is provided for preparing tissue samples. Such a system can be configured for increased throughput during tissue sectioning. The system 100 can be designed to include a block handler 102, one or more microtomes 104, a transport medium 106 (e.g., tape), a hydration chamber 108, and a block tray 110. The block tray 110 can be a drawer-like device designed to hold multiple tissue blocks and can be installed within the system 100 for access by the block handler 102. The block tray 110 can have multiple rows, each designed to hold one or more tissue blocks, and can be sufficiently spaced so that the block handler 102 can deliver, grasp, and remove one tissue block at a time. In some embodiments, the block tray 110 can be designed to securely hold the tissue blocks so that they do not shift or fall out of the block tray 110 during handling, for example, by using a spring-loaded mechanism. In some embodiments, the spring-loaded mechanism can be further designed to enable the block handler 102 to extract tissue blocks without damaging or deforming them. For example, the pitch of the tissue blocks in the block tray 110 can allow the block handler gripper of the block handler 102 to access the paraffin blocks without interfering with adjacent blocks. The block handler 102 can include any combination of mechanisms capable of grasping and / or moving tissue blocks in and out of the microtome 104, specifically into the chuck of the microtome 104. For example, the block handler 102 can include a gantry, push and pull actuators, and grippers on a Selectively Compliant Assembly Robotic Arm (SCARA) robot.

[0041] 1A, in some embodiments, system 100 can include a combination of mechanisms for transferring sections cut from a tissue block onto a transport medium 106 to be transferred to slides for analysis. The combination of mechanisms can include a slide adhesive coater 112, a slide printer 114, a slide input rack 116, a slide singulator that loads slides from a stack of slides 118, and a slide output rack 120. This combination of mechanisms cooperate to prepare samples on slides and to prepare the slides themselves.

[0042] In some embodiments, the one or more microtomes 104 can include any combination of microtomes known in the art, specifically for precisely sectioning tissue blocks. For example, the one or more microtomes 104 can be rotary, cryomicrotome, ultramicrotome, vibrating, saw, laser, etc. based designs. In some embodiments, the one or more microtomes 104 can be designed to move the chuck up and down while also being capable of moving it laterally (e.g., in the direction of the thickness of the block). The one or more microtomes 104 can include any combination of components for receiving and sectioning tissue blocks. For example, the one or more microtomes 104 can include a knife block with a blade handler for holding interchangeable knife blades and a sample holding unit with a chuck head and chuck adapter for holding the tissue block.

[0043] The one or more microtomes 104 are configured to cut tissue sections from a tissue sample encapsulated in a support block of a preservative material, such as paraffin wax. The one or more microtomes 104 can hold a blade that is aligned to cut sections from one face of the tissue block, i.e., the block cutting plane or block face. For example, a rotary microtome can linearly oscillate a chuck that holds the sample block with the cutting surface at the blade cutting plane, which, combined with incremental advancement of the block cutting plane to the cutting plane, allows the microtome 104 to continuously chip thin tissue sections from the block cutting plane.

[0044] In operation, one or more microtomes 104 are used to section and / or slice tissue blocks. When a tissue block is initially delivered to one or more microtomes 104, the tissue block can be sectioned. Sectioning involves removing a layer of protective material to expose a large cross-section of the tissue. That is, the protective material in which the tissue sample is embedded can first be sectioned with relatively thick sections to remove a 0.1 mm to 1 mm layer of paraffin wax above the tissue sample. Once enough paraffin has been removed to expose the complete contours of the tissue sample, the block is "sectioned" and ready to provide processable sections that can be placed on glass slides; the exposed surface is referred to as the block face. Regarding the sectioning process, one or more microtomes 104 can chip away sections of the block until an acceptable portion of the sample within the block is exposed. In some embodiments, the system can include one or more facing cameras to identify when an acceptable portion of the sample within the block is exposed. Regarding the cutting process, one or more microtomes 104 can cut out sample sections of the block with an acceptable thickness to be placed on slides for analysis.

[0045] Once the block is sectioned, in some embodiments, the sectioned tissue block can be hydrated in a hydration fluid for a period of time (e.g., in the hydration chamber 108 or directly in one or more microtomes). In addition to being hydrated, the tissue block can be cooled. The cooling system can be part of the hydration chamber 108 or a separate component from the hydration chamber 108. In some embodiments, the cooling system can provide cooling to all components within the sectioning chamber 150. The sectioning chamber 150 can provide insulation enclosing one or more microtomes 104, the hydration chamber 108, the block tray 110, the blade holder and blade changer of the microtome 104, and the camera. In this way, there are a minimal number of openings in the insulation, which can improve efficiency and effectiveness within the sectioning chamber 150. Regardless of the location, the cooling system can have a mini-compressor, a heat exchanger, and an evaporator plate to create a cool surface. Air within the sectioning chamber can be drawn into and passed across an evaporator plate, for example, using a fan. Cooled air can be circulated within the sectioning chamber 150 and / or hydration chamber 108 to cool the paraffin tissue block. The mass of the equipment within the cooling chamber also provides thermal inertia. Once the chamber is cooled, its temperature can be more effectively maintained if, for example, an access door is opened by the user to remove the block tray 110. In some embodiments, the temperature of the tissue block is maintained between 4°C and 20°C. Keeping the tissue block cool can benefit the sectioning process as well as the hydration process.

[0046] Once the tissue block is sufficiently hydrated, in some embodiments, it is ready for sectioning. Essentially, one or more microtomes cut thin sections of the tissue sample from the tissue block. The tissue sections can then be loaded by a transport medium 106, such as tape, for subsequent transport for placement on slides. In some embodiments, depending on the configuration of the microtomes 104 in the system 100, the system 100 can include a single or multiple transport medium 106 units. For example, in tandem operation, a transport medium 106 can be associated with the polishing and sectioning microtome 104, while in parallel operation, a separate transport medium 106 can be associated with each microtome 104 in the system 100. In an automated system, each of these processes / steps of sectioning, hydration, sectioning, and transfer to slides is computer-controlled rather than being performed in a manual workflow by a histotechnician.

[0047] 1A, 1B, and 1C, in some embodiments, the transport medium 106 can be designed in a manner to which tissue sections cut from a tissue sample in a tissue block can adhere and then be transported by moving the transport medium 106. For example, the transport medium 106 can include any combination of materials designed to physically (e.g., electrostatically) and / or chemically adhere to the sample material. The transport medium 106 can be designed to accommodate multiple tissue sample sections cut from a block to be transferred to a slide for inclusion on the slide for evaluation. In some embodiments, the transport medium 106 can be replaced by a water channel for transporting the tissue. The system 100 can include any additional combination of features for use in automated microtome designs.

[0048] Referring to FIG. 2 , in some embodiments, the system 100 can follow a process 200 for sectioning, hydrating, and sectioning tissue sections and transporting the cut tissue sections to slides in an efficient, automated manner. In some embodiments, each tissue block can have a machine-readable code or identifier thereon. The machine-readable code can include any combination of information for use by the system 100. For example, the machine-readable code can identify the tissue type within the block, such as block 1 having colon tissue, block 2 having tonsil tissue, and block 3 having breast tissue. The information read from the machine-readable code can then be used for other functions. For example, based on the tissue type, the system 100 can determine the hydration time for each tissue type, and block processing can be optimized appropriately. Different tissue types may require hydration for different lengths of time. The hydration time may also depend on the size of the tissue sample. An underhydrated tissue sample may develop microcracks when sectioned. On the other hand, an overly hydrated tissue sample may become overly soft, which may adversely affect cut quality. Thus, in some embodiments, an optimal length of time for which the tissue block should be hydrated can be estimated and used for each tissue block.

[0049] In some embodiments, the system process is provided with a lookup table that can provide hydration times based on prior experiments for appropriate hydration parameters depending on tissue block size or tissue type (or a combination thereof). Such a table can be generated, for example, based on prior experiments with different tissue samples, or can be an estimate based on biophysical / biochemical considerations. In some embodiments, tissue quality can be monitored as a function of hydration parameters, and the table can be updated. Apart from tissue type (which organ), other dependent variables, such as the age of the block or the processing / hydration state of the block, may also need to be considered in generating such empirical data. In some embodiments, data-driven machine learning algorithms can be used to predict optimized parameters for processing, to subsequently optimize such parameters, or both.

[0050] By implementing the process of FIG. 2 , multiple blocks can be processed using overlapping, simultaneous processing steps for the multiple blocks. Generally, an exemplary method includes one or more of steps 250-266. In some embodiments, in step 250, tissue blocks are successively sectioned, starting with block 1, followed by block 2, through block n, and so on. Once sectioned, the tissue blocks are successively hydrated in the same order in which they were sectioned in step 252. In step 254, the amount to which the tissue blocks are hydrated is monitored. Once block 1 is sufficiently hydrated in step 256, it is sectioned in one or more microtomes, and sections from block 1 are loaded with transport media in step 258. In step 260, hydrated block 2, i.e., the next tissue block in the sequence, is sectioned, and sections from block 2 are loaded with transport media in step 262. This process is repeated until block n is hydrated in step 262, sectioned in step 264, and the sections from block n are loaded with a transport medium in step 264.

[0051] As a non-limiting example, the process of FIG. 2 will be described in more detail. First, a user can place one or more tissue blocks on a block tray 110 to be loaded into the system 100. In some embodiments, once the block tray 110 is in position, a scanning device (e.g., a barcode scanning device) attached to the block handler 102 can scan one or more machine-readable codes (e.g., barcodes) printed on the paraffin tissue blocks in the block tray 110. The block tray 110 can be designed to expose these machine-readable codes to a scanning device or reader for efficient identification. In some embodiments, the block handler 102 can load the tissue blocks and position them in proximity to a fixed scanning device to be identified. The block tray 110 and the sample chucks on one or more microtomes 104 can be oriented so that the block handler 102 has a minimum number of degrees of freedom, which saves cost and keeps its design simple. This can also reduce the number of movements each time a tissue block is transferred between the chuck and the block tray 110, which can save time. Once the tissue block has been identified, the block handler 102 can place the block on the microtome 104 for processing.

[0052] One or more microtomes 104 can then section the first tissue block using any combination of sectioning methods. System 100 can section multiple blocks one at a time and place them sequentially in hydration chambers, thereby allowing for batch sample processing. Block handler 102 can move the first sectioned tissue block into hydration chamber 108 (although in some embodiments, hydration can occur in one or more microtomes). A processor within system 100 can then track the hydration time of the first tissue block (and each subsequent tissue block) within hydration chamber 108. The next tissue block can be similarly sectioned and hydrated. Once a predetermined period of time has elapsed (once the block is properly hydrated), block handler 102 can move the first sectioned tissue block from hydration chamber 108 into the chuck of one or more microtomes 104. The first tissue block can be sectioned while other blocks, which begin hydrating thereafter in a sequential manner, continue to hydrate. As one or more microtomes 104 cut one or more sections from the sectioned portion of the first tissue block, the cut sections can be transferred to the transport medium 106. Then, the second tissue block can be hydrated and sectioned, followed by the next tissue block. The steps are repeated until all tissue blocks from the block tray 110 have been processed.

[0053] 2 illustrates the automated steps in batch processing, as it depicts the sequential sectioning of blocks (block 1...n), the sequential movement of the blocks into the hydration chamber 108 within the system 100, and the movement from the hydration chamber 108 to the chuck of one or more microtomes 104 in a first-in, first-out process (responsive to automated timing calculations) for sectioning by the microtome 104. As can be appreciated, this first-in, first-out method can improve the flow and efficiency of the system 100 because hydrated blocks can be sectioned while other blocks are being hydrated. In some embodiments, one or more block handlers 102 can be programmed to move various blocks around throughout the process 200 to maintain the most efficient flow. For example, the block handler 102 can move sectioned blocks to the hydration chamber 108 and back to the chuck of the microtome 104 for tissue sectioning according to a preprogrammed timer that is timed according to the period required for hydration.

[0054] 2, system 100 can process multiple blocks in a batch format with overlapping, simultaneous processing steps for multiple blocks, rather than in a fully sequential format requiring each block to be fully processed before processing of the next block begins. In some embodiments, batch sectioning can be performed as a FIFO (first in, first out) queue, such that a series of blocks are placed in hydration (e.g., an ice-water mixture) for a predetermined period of time. Once a first tissue block is sufficiently hydrated, the tissue block is ready for abrasive cutting, and sections can be taken (cut) from this first tissue block. Meanwhile, other blocks continue to hydrate.

[0055] However, it should be noted that in some embodiments, a continuous workflow or a combination of continuous workflow and batch processing may be utilized. Figure 3 illustrates the difference between batch processing and continuous workflow. The diagram in Figure 3 compares a batch workflow with a continuous workflow in an example showing the processing of three tissue blocks. The vertical axis 301 of the diagram in Figure 3 represents elapsed time, with blocks to the left of the vertical black line representing sample flow in batch block processing and blocks to the right of the vertical black line representing sample flow in continuous processing. In a continuous process, a first tissue block is sectioned by the microtome 104 and then placed in the hydration chamber 108 for the required period of time. Once hydration is complete, the automated system 100 will continue by removing the tissue block, placing it on the chuck of the microtome 104, and cutting tissue sections from the tissue block for transfer of the cut tissue sections to a transport medium 106, such as tape, within the automated device.

[0056] In some embodiments, batch processing can be performed by a single microtome 104 or multiple microtomes working in parallel or tandem. One or more microtomes 104 can continue cutting tissue blocks while other tissue blocks (uncut samples) are hydrating to avoid time delays. As shown in FIG. 3 , on the left, block 1, block 2, and block 3 can each be in a particular step of process 200 at the same time. In some embodiments, a single block handler 102 can be responsible for managing all of the movement and placement of tissue blocks, so the process flow may have staggered starts and finishes between adjacent blocks. It should be understood that the columns in FIG. 3 depict time increments; thus, block sectioning takes a first period (e.g., 1 increment), sectioning takes a longer period (e.g., 2 increments, i.e., twice the amount of time), while hydration takes the longest period (e.g., 31 increments). In parallel batch processing, all three blocks are sectioned, at least some of them, while only one block is sectioned during sequential processing in the same amount of time. This increased number of time increments in sequential processing is depicted in the graph. This contrasts with the conventional process depicted on the right, where a complete process step is completed prior to beginning any of the steps for a subsequent block. Note that the disclosed embodiments involving multiple microtomes can be utilized in both batch processing and sequential workflows.

[0057] Note that the time increments on the graph are relative (dimensionless) and do not depict defined periods of time. The time increments are shown as examples because the relative times for hydration, sectioning, and sectioning may differ from those shown; however, it is further noted that in any event, the hydration time is a multiple of the sectioning time, and therefore batch processing in an automated device would reduce the overall time, thereby improving (increasing) the throughput of the system. Note that the graph shows three blocks to illustrate a comparison of when additional blocks would be sectioned in a typical process.

[0058] 1A, 1B, and 1C, in some embodiments, the system 100 can include multiple microtomes 104 operating within the system 100 to increase the processing capacity of the automated system 100. While the system will be described with reference to two microtomes to aid in understanding the present disclosure, it should be noted that any number of microtomes can be used in accordance with the present disclosure, and the number of microtomes can be even or odd. All of the microtomes 104 can be used in an automated manner (e.g., controlled by a processor or controller) to process (e.g., section and cut) tissue blocks in an efficient manner. In some embodiments, as discussed with respect to FIGS. 4A and 4B, some of the microtomes 104 can be used to section all of the tissue blocks, while other microtomes 104 can work in tandem to cut the sectioned tissue blocks. In some embodiments, as discussed with respect to Figures 5A and 5B, multiple microtomes 104 can each be used to both section and cut individual tissue blocks, while other microtomes 104 can work in parallel to section and cut other individual tissue blocks. In some embodiments, the blade holders for each microtome 104 are referenced to one another to reduce the number of abrasive cuts. In some embodiments, the microtomes 104 can be located on the same base plate with machined hole patterns that will orient the microtomes 104 relative to one another in a predetermined orientation. Each blade holder and chuck on the microtome 104 can also be referenced to the microtome structure through machined holes and fasteners. Using machined holes can reference the two microtomes very closely, but not perfectly, to one another. Final referencing can be achieved through abrasive cuts on the sectioning microtome 104. The abrasive cut can align the tissue block surface so that it faces the sectioning microtome 104 on the sectioning microtome 104 .In some embodiments, system 100 can be designed to reduce or substantially eliminate vibrations between microtomes 104 so that they do not adversely affect each other's cut quality. For example, vibrations can be passively controlled by having a large mass around the moving microtome parts and a softer connection to the common base. In this way, the microtome can act like a rigid body. The critical path for vibration on a given microtome is between the blade and the paraffin block; therefore, system 100 can be designed to minimize the number of connections and fasteners between the two endpoints to create a rigid system and control vibrations.

[0059] In some embodiments, some of the microtomes may be dedicated to sectioning the block, while others may be dedicated to sectioning the block. After an initial set of block hydration, when the FIFO queue becomes saturated, multiple microtomes will work in tandem to complete the work. In an alternative workflow embodiment, multiple microtomes can be utilized for sectioning and sectioning. In this way, the microtomes will be utilized 100% of the time. This accelerates the workflow by providing multiple paths (redundancy) for the same task. The device / system can have a smart decision algorithm for which workflow to use and when.

[0060] 4A and 4B, in some embodiments, two or more microtomes 104 can be configured in system 100 to operate in tandem or serially. Figures 4A and 4B depict an exemplary process 400 for operating system 100 with two or more microtomes 104 operating in tandem. Figures 4A-4B include various steps 402-450, some of which are optional. Some of the steps are discussed in more detail below.

[0061] First, a user can place one or more tissue blocks on the block tray 110 to be loaded into the system 100. In some embodiments, the block handler 102 can have specialized grippers designed to load tissue blocks and move them between the various locations discussed in FIGS. 4A and 4B . For example, the block handler 102 can have specialized grippers designed to move blocks from the block tray 110 to the chuck of the sectioning microtome 104, and then to the hydration chamber 108, and then back to the sectioning microtome 104. The grippers can have finger-like features that open and close, and with this action, the grippers can hold the blocks and apply sufficient force to safely remove them, for example, from the chuck on the microtome 104. In some embodiments, the hydration chamber 108 has a structure similar to the block tray 110 and can hold the tissue blocks in place while allowing the block handler 102 to easily place and remove blocks. Supervision software can be provided to supervise and coordinate the actions of the microtome 104, block handler 102, and hydration chamber 108.

[0062] First, in some embodiments, once the block tray 110 is in place, a barcode scanning device attached to the block handler 102 can scan one or more barcodes printed on the paraffin tissue blocks in the block tray 110. In some embodiments, the block handler 102 can load the tissue blocks and position them in proximity to the fixed barcode device for identification.

[0063] Once the tissue block is identified in step 402, the block handler 102 can transport the first tissue block (block n) to one of the microtomes 104 for sectioning. For example, the block handler 102 can reposition the tissue block to a microtome 104 designated as the sectioning microtome 104. In some instances, the block handler 102 can remove the first tissue block from the block tray 110 and move it toward one of the microtomes 104 for sectioning.

[0064] In some embodiments, a supervision algorithm can use machine-readable codes on the blocks to direct where the block handler 102 transports and places the blocks. For example, the block handler 102 can include a scanning device to read the machine-readable codes on the blocks before loading them. In this way, the supervision algorithm can identify and track the blocks, providing instructions to the block handler 102 regarding which blocks to load and where to place them.

[0065] In step 404, the block handler 102 can place a first tissue block into the sectioning microtome 104 (e.g., in the chuck). In some embodiments, the chuck of the sectioning microtome 104 can move up and back in preparation to receive a new block. Following or simultaneously with preparing the microtome 104, the block handler 102 can load a block from the block tray 110, bring it into the chuck of the sectioning microtome 104, and slide it from right to left. In some embodiments, the chuck can have a spring-loaded clamping mechanism, and the gripper of the block handler 102 can have sufficient strength to overcome this clamping force. Once the block is inserted into the chuck, the gripper of the block handler 102 releases its finger-like features and moves slightly toward the blade holder, disengaging the block. The block is now securely held by the microtome chuck.

[0066] In step 406, with the first tissue block in place within the chuck, the sectioning microtome 104 can begin the sectioning process and remove any excess paraffin layer on the tissue resulting from the molding process. In some embodiments, the microtome can move the microtome chuck to a position sensor, which can be a touch sensor or a non-touch laser or sound-based sensor. Each block can be fabricated in a mold that can have a slightly different height. The system 100 needs to know the location of the surface of the first tissue block relative to the plastic cassette. This sensor can determine the location of the block's surface. For a touch sensor, a resistive sensor can be used that changes a voltage output sensed by control hardware. A change in voltage indicates that the block surface is touching the sensor. If the block thickness axis encoder is read simultaneously, this gives the location of the block surface. For a non-touch laser implementation, a through-beam laser sensor can be used. There would be an emitter and receiver that are spaced apart from each other but on the same line of sight. As the block is advanced along the thickness axis (x-axis), it interrupts the optical path, and a change in the voltage reading at the receiver can be observed. This voltage change and the encoder reading along the thickness axis simultaneously indicate the location of the block surface. Regardless of the type of sensor used, the surface detection sensor can be placed at a known location relative to the microtome blade. After surface detection, the chuck can be moved to the sectioning position as close as possible to the blade in the vertical plane. After several up-and-down microtome cycles, the surface of the first tissue block will catch on the blade.

[0067] In step 408, in some embodiments, a strategically placed camera can be provided to capture images of the first tissue block as it is being cut. The camera can be designed to provide image data for determining when sufficient sections have been taken and a large cross-section of the tissue is exposed. During the sectioning process, paraffin chips are created as waste. In some embodiments, these paraffin chips can be blown off by an air blade on the chuck and sucked up by a vacuum on the blade holder.

[0068] In step 410, the block handler 102 can remove the first tissue block from the sectioning microtome 104 and place the first tissue block in the hydration chamber 108. Removing the block from the sectioning microtome 104 can be the reverse of placing the block on the chuck from step 406, but with the destination being the hydration chamber 108.

[0069] In step 412, the first tissue block hydrates in the hydration chamber 108 for a predetermined period of time. For example, the block can be designed to spend approximately 10-15 minutes in the hydration chamber 108. A supervisory control algorithm can command the block handler to load the block with the longest hydration time first to be sectioned first. In some embodiments, sensors can be used to detect the hydration level of the tissue. For example, the hydration level of the block can be periodically measured so that once a predetermined hydration level is reached, the block can be marked for removal for the next step. In some embodiments, the hydration time for each tissue type can be appropriately optimized to account for the tissue type in the block. For example, a block containing colon tissue may have a different hydration level / time than a block containing breast tissue.

[0070] In step 414, subsequent to or substantially simultaneous with step 410, the block handler 102 may move a second tissue block (block k) to the sectioning microtome 104. Steps 414-424 are substantially identical to steps 402-412 performed on the first tissue block, but may be performed on the second tissue block. In this mode of operation, the sectioning microtome sections the first tissue block, and while the first tissue block is hydrating, the sectioning microtome begins sectioning the second tissue block.

[0071] In step 432, once the first tissue block is properly hydrated, the block handler 102 can transport the first tissue block to one of the microtomes 104 for polishing and sectioning. For example, the block handler 102 can reposition the tissue block to a microtome 104 designated as the polishing and sectioning microtome 104.

[0072] In step 434, the block handler 102 can place the first tissue block into (e.g., in a chuck) the polishing and sectioning microtome 104, for example, using a similar process as discussed in step 406.

[0073] In step 436, with the first tissue block in place within the chuck, the polishing and sectioning microtome 104, in some embodiments, moves the microtome chuck to a sensor that can identify the surface of the first tissue block with exposed tissue (e.g., the sectioned portion of the block). For example, a line scan camera can be used to observe the paraffin removal from the block. When the camera detects that the tissue width is constant throughout the length of the block, it is considered fully polished and flattened on the sectioning microtome blade. Because the polishing and sectioning microtome 104 and the sectioning microtome 104 are different instances of a microtome 104, due to the manufacturing / assembly process, they are not identical on the 4 μm scale (4 μm is the tissue section thickness). The sectioning cuts can be 10-15 μm thick, and the polishing cuts can be 4-5 μm thick.

[0074] In step 438, once the surface of the tissue block is found, the first tissue block is positioned on the cutting blade and an abrasive cut is made. The abrasive cut has the same thickness as the final tissue section, but is not transferred to a glass slide. The abrasive cut is required to ensure that the first tissue block surface and the blade are in substantially the same vertical plane. The abrasive cut creates a plane on the surface of the block so that the blade remains on that plane throughout the height of the block. Once the abrasive cut is completed, an automated device applies tape to the first tissue block surface. An adhesive transport medium 106 is present, deployed from the blade holder, and the chuck of the polishing and sectioning microtome 104 gently presses the paraffin block surface against the adhesive side of the transport medium 106. In some embodiments, the rear wall of the transport medium 106 can support the transport medium 106 as it is pressed against the paraffin block and the chuck of the polishing and sectioning microtome 104. In some embodiments, there is a resistive sensor behind the transport medium that picks up a signal when the block is pressed against the transport medium. This indicates that at least a certain point on the block has touched the transport medium. Similar but more complex sensors can be used to perform pressure mapping. In this case, the percentage of the block face that is touching the transport medium can be determined. Once the transport medium 106 is attached, it can perform a cut and the first tissue block can be divided into two distinct parts: the one attached to the plastic cassette (the remaining block) and the tissue section.

[0075] In step 440, the block handler 102 may remove the first tissue block from the polishing and sectioning microtome 104 via the transport medium 106. For example, the tissue section may be attached to tape and transported to another part of the system 100. At this point, the processing cycle for the first tissue block has completed the tandem process 400.

[0076] In step 442, subsequent to or substantially simultaneous with step 440, the block handler 102 can move a second tissue block to the polishing and sectioning microtome 104. Steps 442-450 can be substantially identical to steps 432-440 performed on the first tissue block, but can be performed on the second tissue block.

[0077] Continuing with the tandem process 400 of FIGS. 4A and 4B, all blocks will proceed through both the sectioning microtome 104 and the polishing and sectioning microtome 104. One advantage of using multiple microtomes 104 in tandem would be that other support subsystems would not need to be duplicated. For example, the transport medium 106 handler subsystem is a large and complex system; in tandem operating mode, one transport medium 106 handler would be sufficient to handle the blocks processed by the two microtomes 104. Also, in tandem operating mode, any combination of sensors optimized for their task can be installed on each microtome 104. For example, the sectioning microtome could have a camera with UV light and a visible camera or a mid-wave infrared (MWIR) LED and camera, while the sectioning microtome could have a visible camera and a pressure sensor. The steps of FIGS. 4A and 4B improve the performance and quality of the work output.

[0078] 5A and 5B, in some embodiments, two or more microtomes 104 can be configured in system 100 to operate in parallel. Figures 5A-5B include various steps 502-564, some of which are optional. Some of the steps are discussed in more detail below.

[0079] For example, system 100 can be designed so that multiple microtomes 104 can operate in parallel to section, polish, section, and tape samples from a block. In this workflow, a given block is processed by only some, but not all, of the microtomes, for example, according to process 200 discussed with respect to FIG. 2. In some embodiments, as shown in FIGS. 5A and 5B, each of the microtomes 104 operating in parallel can be phased in start time so that they can share subsystems (e.g., mounting mechanism, hydration chamber 108, transport medium 106 handler, etc.). Process 500 in FIGS. 5A and 5B includes the same steps provided in FIGS. 4A and 4B, but each microtome 104 operating in parallel can perform the steps associated with the sectioning microtome 104 and the polishing and sectioning microtome 104, respectively. For example, one or more first microtomes 104 would section a first tissue block, which would then be returned to the first microtome 104 after polishing and hydration for sectioning. Substantially parallel to the processing of the first tissue block by the first microtome 104, one or more second microtomes 104 would section a second tissue block, which would then be returned to the second microtome 104 after polishing and hydration for sectioning. This process 500 can reduce the time required to resurface the block after hydration. In some embodiments, the microtomes 104 can be the same, reducing the number of parts involved from a manufacturing perspective. It is also flexible and can receive input from a user to select an internal device flow that will better function for the laboratory's overall workflow.

[0080] Continuing with FIGS. 5A and 5B , a user can first place one or more tissue blocks on the block tray 110 to be loaded into the system 100. In some embodiments, the block handler 102 can have specialized grippers designed to load tissue blocks and move them between the various locations discussed in FIGS. 5A and 5B . For example, the block handler 102 can have specialized grippers designed to move blocks from the block tray 110 to the chuck of the first microtome 104, and then to the hydration chamber 108, and then to the sectioning microtome 104. The grippers can have finger-like features that open and close, and with this action, the grippers can hold the block and apply sufficient force to safely remove it, for example, from the chuck on the microtome 104. In some embodiments, the hydration chamber 108 has a structure similar to the block tray 110 and can hold the tissue block in place while allowing the block handler 102 to easily place and remove blocks. Supervision software can be provided to supervise and coordinate the actions of the microtome 104, block handler 102, and hydration chamber 108.

[0081] First, in some embodiments, once the block tray 110 is in place, a barcode scanning device attached to the block handler 102 can scan one or more barcodes printed on the paraffin tissue blocks in the block tray 110. In some embodiments, the block handler 102 can load the tissue blocks and position them in proximity to the fixed barcode device for identification.

[0082] Once the tissue block is identified in step 502, the block handler 102 can transport the first tissue block (block n) to one of the microtomes 104 for sectioning. For example, the block handler 102 can reposition the tissue block to a microtome 104 designated as the first microtome 104. In some instances, the block handler 102 can remove the first tissue block from the block tray 110 and move it toward one of the microtomes 104 for sectioning.

[0083] In some embodiments, a supervision algorithm can use machine-readable codes on the blocks to direct where the block handler 102 transports and places the blocks. For example, the block handler 102 can include a scanning device to read the machine-readable codes on the blocks before loading them. In this way, the supervision algorithm can identify and track the blocks, providing instructions to the block handler 102 regarding which blocks to load and where to place them.

[0084] In step 504, the block handler 102 can place a first tissue block into the first microtome 104 (e.g., in the chuck). In some embodiments, the chuck of the first microtome 104 can move up and back in preparation for receiving a new block. Following or simultaneously with preparing the microtome 104, the block handler 102 can load a block from the block tray 110, bring it into the chuck of the first microtome 104, and slide it from right to left. In some embodiments, the chuck can have a spring-loaded clamping mechanism, and the gripper of the block handler 102 can have sufficient strength to overcome this clamping force. Once the block is inserted into the chuck, the gripper of the block handler 102 releases its finger-like features and moves slightly toward the blade holder, disengaging the block. The block is now securely held by the microtome chuck.

[0085] In step 506, with the first tissue block in place within the chuck, the first microtome 104 can begin the sectioning process and remove any excess paraffin layer on the tissue resulting from the molding process. In some embodiments, the microtome can move the microtome chuck to a position sensor, which can be a touch sensor or a non-touch laser or sound-based sensor. Each block can be fabricated in a mold that can have a slightly different height. The system 100 needs to know the location of the surface of the first tissue block relative to the plastic cassette. This sensor can determine the location of the block's surface. For a touch sensor, a resistive sensor can be used that changes a voltage output sensed by control hardware. A change in voltage indicates that the block surface is touching the sensor. If the block thickness axis encoder is read simultaneously, this provides the location of the block surface. For a non-touch laser implementation, a through-beam laser sensor can be used. There would be an emitter and receiver spaced apart from each other but on the same line of sight. As the block is advanced along the thickness axis (x-axis), it interrupts the optical path, and a change in the voltage reading at the receiver can be observed. This voltage change and the encoder reading along the thickness axis simultaneously indicate the location of the block surface. Regardless of the type of sensor used, the surface detection sensor can be placed at a known location relative to the microtome blade. After surface detection, the chuck can be moved to the sectioning position as close as possible to the blade in the vertical plane. For example, after several up-and-down microtome cycles, the surface of the first tissue block catches on the blade.

[0086] In step 508, in some embodiments, a camera can be provided to capture images of the first tissue block as it is being cut. The camera can be designed to provide image data for determining when sufficient sections have been taken and a large cross-section of the tissue is exposed. During the sectioning process, paraffin chips are created as waste. In some embodiments, these paraffin chips can be blown off by an air blade on the chuck and sucked up by a vacuum on the blade holder.

[0087] In step 510, the block handler 102 can remove the first tissue block from the first microtome 104 and place the first tissue block in the hydration chamber 108. Removing the block from the first microtome 104 can be the reverse of placing the block on the chuck from step 406, but with the destination being the hydration chamber 108.

[0088] In step 512, the first tissue block hydrates in the hydration chamber 108 for a predetermined period of time. For example, the block can be designed to spend approximately 10-15 minutes in the hydration chamber 108. A supervisory control algorithm can command the block handler to load the block with the longest hydration time first to be sectioned first. In some embodiments, sensors can be used to detect the hydration level of the tissue. For example, the hydration level of the block can be periodically measured so that once a predetermined hydration level is reached, the block can be marked for removal for the next step. In some embodiments, the hydration time for each tissue type can be appropriately optimized to account for the tissue type in the block. For example, a block containing colon tissue may have a different hydration level / time than a block containing breast tissue.

[0089] In step 514, following or substantially simultaneously with step 502, the block handler 102 can move a second tissue block (block m) to the second microtome 104. In some embodiments, supervisory level software can track where each block is sectioned so that it can be returned to the same microtome after hydration. Steps 514-524 can be substantially identical to steps 502-512 performed on the first tissue block by the first microtome 104, but performed on the second tissue block by the second microtome 104. In this mode of operation, the first microtome first sectioned the first tissue block, and while the first tissue block was hydrating, the first microtome began sectioning the second tissue block.

[0090] In step 526, following or substantially simultaneously with step 514, the block handler 102 may move a third block (block k) to the first microtome 104. In some embodiments, supervisory level software may track where each block is sectioned so that it can be returned to the same microtome after hydration. Steps 526-536 may be substantially identical to steps 502-512 performed by the first microtome 104 on the first tissue block, but performed by the first microtome 104 on the third block. In this mode of operation, the first microtome will section the third tissue block while the first tissue block is hydrating.

[0091] In step 538, following or substantially simultaneously with step 526, the block handler 102 may move a fourth block (block i) to the second microtome 104. In some embodiments, supervisory level software may track where each block is sectioned so that it can be returned to the same microtome after hydration. Steps 538-548 may be substantially identical to steps 502-512 performed on the first tissue block by the first microtome 104, but performed on the fourth block by the second microtome 104. In this mode of operation, the second microtome will section the fourth tissue block while the second tissue block is hydrating.

[0092] In step 550, once the first tissue block is properly hydrated, the block handler 102 can transport the first tissue block to the first microtome 104 for polishing and sectioning. In step 552, the block handler 102 can place the first tissue block into the first microtome 104 (e.g., in a chuck). In step 554, with the first tissue block in place in the chuck, the first microtome 104, in some embodiments, can move the microtome chuck to a sensor to identify the surface of the first tissue block with exposed tissue (e.g., the sectioned portion of the block). For example, a line scan camera can be used to observe the paraffin removal from the block. When the camera detects that the tissue width is constant throughout the length of the block, it is considered fully polished and flattened on the sectioning microtome blade.

[0093] In step 556, once the abrasive cutting is completed, the automated device applies tape to the first tissue block face. There is an adhesive transport medium 106 deployed from the blade holder, and the chuck of the first microtome 104 gently presses the paraffin block face against the adhesive side of the transport medium 106. In some embodiments, a rear wall of the transport medium 106 can support the transport medium 106 as it is pressed against the paraffin block and the chuck of the second microtome 104. In some embodiments, there is a resistive sensor behind the transport medium that picks up a signal when the block is pressed against the transport medium. This indicates that at least a point on the block has touched the transport medium. Similar but more complex sensors can be used to perform pressure mapping. In this case, the percentage of the block face that is touching the transport medium can be determined. Once the transport medium 106 is attached, it can perform a cut, and the first tissue block can be divided into two distinct parts: the part attached to the plastic cassette (the remaining block) and the tissue section. The block handler 102 may also remove the first tissue block from the first microtome 104 via the transport medium 106. For example, the tissue section may be taped and transported to another part of the system 100. At this point, the processing cycle for the first tissue block has completed the parallel process 400.

[0094] In step 558, following or substantially simultaneously with step 550, the block handler 102 can move the second tissue block to the second microtome 104. Steps 558-564 can be substantially identical to steps 550-556 performed on the first tissue block, but performed on the second tissue block. Similarly, the third and fourth blocks can be polished and taped using similar processing steps. Continuing with the parallel process 500 of Figures 5A and 5B, all blocks will proceed through both the first microtome 104 and the second microtome 104. The steps of Figures 5A and 5B improve performance and quality of the work output.

[0095] It should be noted that although the above process is discussed in relation to moving the tissue block to a hydration chamber, in some embodiments, the hydration of the tissue block may be implemented in place in one or more microtomes.

[0096] Figures 6, 7A, and 7B, as an example, depict charts 600, 700, 800 showing comparative block hydration capacity over time for 75 tissue blocks, with time increments assigned numerical values ​​for ease of illustration. Figure 6 illustrates block hydration capacity using a single microtome, while Figures 7A and 7B illustrate block hydration capacity using two microtomes in accordance with the present disclosure. Figure 7A depicts a tandem operating mode, the lower graph depicts a parallel operating mode, and Figure 7B depicts a parallel operating mode. Note that 75 blocks are shown as an example to illustrate the advantage of additional microtomes.

[0097] As the chart shows, in this example, device throughput can be increased by more than 18% under certain run conditions by using a second microtome. The X-axis provides dimensionless time, and the Y-axis provides the number of blocks (block count). Line A (dotted line) designates the number of blocks sectioned, line B (dashed line) designates the number of blocks hydrating, and line C (dash-dotted line) designates the number of blocks sectioned. As can be seen, over a period of time, blocks are sectioned and placed in a mixture or chamber through the hydration phase. After the first tissue block is hydrated, sectioning can begin using the second microtome, while the first microtome continues to section the block. As blocks continue to be removed from the hydration phase (lowering of the hydration line until all blocks are sectioned) and blocks continue to be sectioned to completion, the time unit in Figures 7A and 7B is at 146, while the time unit in Figure 6 is at 176. Also note that the slope of the lines in Figures 7A and 7B is constant, while the lines in Figure 6 are zigzag, illustrating that the average speed is slower in Figure 6. A comparison of Figure 6 and Figures 7A and 7B illustrates the increased throughput using two microtomes in accordance with one embodiment of the multiple-pass process of the present disclosure.

[0098] It should be understood that the graphs in Figures 6, 7A, and 7B are provided to illustrate examples of the benefits of using multiple microtomes, and that other block counts and other time periods are also contemplated. Additionally, if more than two microtomes are used in an automated system, the throughput will increase by amounts far beyond those depicted in Figures 7A and 7B. Thus, it should be understood that in some embodiments in which more than two microtomes are utilized in the system for block sectioning and / or tissue sectioning, the processing speed is increased. In some embodiments involving multiple microtomes, a combination of parallel, tandem, sequential, or even batch methods can be used.

[0099] To further increase throughput with automated tissue sectioning equipment, the present disclosure further provides improved hydration methods. In some embodiments, the disclosed methods enhance the rate of hydration by overcoming certain fundamental aspects of the wax medium, particularly its hydrophobicity, as evidenced by its high contact angle with the hydrating liquid. For example, the methods can employ surfactants to reduce the contact angle of the hydrating liquid on hydrophobic surfaces. In addition, surfactants can also enable the hydrating liquid to diffuse into the biological tissue material at a much faster rate. It is also highly desirable to further promote capillary infiltration of the hydrating liquid into the paraffin pores, separate from more hydrophilic biological tissue materials, such as carbohydrates, proteins, and lipids. In some embodiments, the present disclosure provides methods for either supplying or transporting the hydrating liquid, used alone or in conjunction with the aforementioned enabling surfactants, to further improve the efficiency of hydration.

[0100] In some embodiments, a hydrating liquid vapor can be applied onto the tissue block in the hydration chamber such that hydrating liquid droplets condense on the block face (the exposed surface of the tissue sample). In some embodiments, a directional flow of moisture to the block face is applied. In some embodiments, a directional flow of hydrating liquid droplets to the block face can be applied through a spray application, such as by a sprayer. In some embodiments, the disclosed methods allow for hydration of single or multiple blocks to occur either alone or in a dedicated hydration chamber, which may further lend itself to the use of several additional classes of surfactants to be used in conjunction with the hydrating liquid in any form.

[0101] Typically, as described above, tissue samples are provided as tissue blocks or sample blocks in which the tissue is embedded in a preservative material such as paraffin. The tissue block is then sectioned by removing the layer of preservative material in which the tissue is embedded, exposing a large cross-section of the tissue; such exposed section of the tissue sample is referred to as the block face. Once the tissue block is sectioned, the tissue sample in the tissue block can be hydrated. In some embodiments of the present disclosure, higher throughput is achieved by novel methods and systems for accelerated tissue / wax hydration. For example, sectioning a block typically takes less than one minute, while hydration and cooling can take up to 30 minutes. According to current pathology laboratory practice, unless the block is properly hydrated and cooled, it cannot be mounted on a glass slide. Therefore, by accelerating the hydration step, the overall process of tissue sampling can be significantly expedited.

[0102] In some aspects of the present disclosure, tissue blocks are hydrated by condensing hydrating liquid droplets onto the block face to facilitate sectioning in a microtome. Other chemical additives, separate from the pure hydrating liquid, either embodied within the condensed liquid droplets or otherwise applied to the block face in a prior step, may also be used. In some embodiments, such chemicals may include, but are not limited to, oil-soluble surfactants applied directly to the block face to induce rapid hydrating liquid penetration into the wax, resulting from the emulsification of the rapidly condensing hydrating liquid droplets from the air saturated with hydrating liquid vapor (such air is also synonymously referred to as humid air, although it should be noted that hydrating liquids other than water may also be used). In some embodiments, such chemicals may be soluble in the hydrating liquid, thereby enhancing the spreading and penetration of the liquid droplets. Liquid droplets can be applied, for example, as a spray onto the block face or by direct droplet condensation of a (diluted) surfactant solution in a humidified chamber.

[0103] According to some aspects of the present disclosure, tissue blocks are placed in a sealed hydration chamber for a defined period of time, and droplet condensation on the block surface allows for very rapid hydration to occur. In some embodiments, droplet condensation occurs under very controlled conditions in a dedicated hydration chamber, thus allowing for batch processing of many tissue blocks simultaneously.

[0104] According to some aspects of the present disclosure, a hydration chamber generates condensed hydrated liquid droplets on a cold block surface, and the warmer temperature of the condensing hydrated liquid droplets not only enhances the kinetic energy of the impacting droplets, but the resulting heat of condensation also softens the wax in close proximity to the air / tissue interface. In this way, hydration liquid entrainment is kinetically accelerated, effectively reducing tissue hydration time and therefore improving device productivity of the automated tissue sectioning apparatus.

[0105] According to some aspects of the present disclosure, hydration methods can also include directing a stream of moisture onto the block face (e.g., using a sonicator) and may also include the application of hydrating liquid droplets on the block face. The directed spray of hydrating liquid, either in the vapor phase as moisture or in the liquid phase as condensed hydrating liquid droplets, can be variously derived or generated forms of hydrating liquid that serve to hydrate individual block faces or multiple tissue blocks. In some embodiments, such processes may not require a separate hydration unit, but humidification or spraying may instead be achieved in the microtome. In some embodiments, this may be achieved by a dispersed spray of hydrating liquid droplets, or alternatively, a directed or focused spray of moisture, which may subsequently condense on a cool block face maintained below the dew point of the hydrating liquid vapor.

[0106] According to some aspects of the present disclosure, a chemical additive can be applied to the block face, which further facilitates or accelerates the process of aqueous diffusion into the tissue / wax composite medium. The block face may then undergo any form of humidification, including both liquid (hydrating liquid droplets) and gas (water that tends to condense on the cold block face), such as a directed spray of hydrating liquid droplets, a focused stream of moisture (hydrating liquid vapor), or any other method that can serve as a source of hydrating liquid. Consistent with the specific (thermodynamic) activity of the chemical additive, aqueous diffusion can thereby be accelerated, and the enhanced rate of tissue hydration may result from any number of mechanisms involving various physical or chemical forces, such as surface tension, diffusion, and wetting, as non-limiting examples, that allow the hydrating liquid to overcome the transport resistance associated with the hydrophobic properties of the wax, i.e., its high contact angle with the hydrating liquid.

[0107] According to some aspects of the present disclosure, an oil-soluble surfactant applied directly to the block surface induces very rapid hydration liquid penetration into the wax resulting from the emulsification of rapidly condensing hydration liquid droplets from moist air. In this hydration liquid / oil (W / O) emulsification process, moist air is maintained such that hydration liquid droplets condense onto the very cool, surfactant-coated block surface, and hydration of paraffin-infused tissue results from the thermodynamic driving force for the (instantaneous) emulsification of the condensed hydration liquid vapor droplets on the surfactant-stabilized wax surface.

[0108] According to some aspects of the present disclosure, the hydration method also includes directing a stream of water (e.g., using a sonicator) onto the block face to which the surfactant additive was previously applied. A directed spray of hydration liquid, either in the gas phase as water or in the liquid phase as a spray of hydration liquid droplets, is then used to hydrate the tissue block. Such a process may not necessarily require a separate humidification unit and may be performed directly in the microtome.

[0109] According to some aspects of the present disclosure, a super-spreading aqueous surfactant solution is sprayed directly onto the block face or otherwise condensed as liquid microdroplets onto the block face from within a hydration chamber prior to sectioning in a microtome. The deposited or condensed aqueous solution spreads very rapidly across the tissue / wax surface, simultaneously allowing very rapid penetration of the hydrating liquid into the tissue therein exposed to the block face.

[0110] Once hydration is complete, the blocks are sectioned and transferred to adhesive tape for further downstream processing.

[0111] Referring to FIG. 8 , an exemplary method for an enhanced hydration process is described. Generally, the exemplary method includes one or more of steps 810-824. In some embodiments, in step 810, a sectioned tissue block is placed in a humidification chamber. In step 812, the tissue block can be hydrated, which in some embodiments involves hydrating the tissue block by hydration liquid vapor condensation in step 814. In some embodiments, in step 816, alternatively or additionally, the tissue block is hydrated by directed spraying of hydration liquid droplets. In step 818, an optional surfactant is used. In some embodiments, in step 820, an oil-based surfactant is used. In some embodiments, in step 822, alternatively or additionally, a super-diffusing surfactant can be used. In step 824, the tissue block is sectioned in a microtome.

[0112] In some embodiments, in step 814, the tissue block can be placed in a hydration chamber for a defined time, and droplet condensation on the sectioned block allows for very fast hydration. In some embodiments, the tissue block can be hydrated using rapidly condensing hydrating liquid droplets applied to the block face. In some embodiments, this may be achieved using a sonicator. In some embodiments, humid air can be maintained in the hydration chamber such that the hydrating liquid droplets condense onto a very cool surfactant-coated block face (approximately 4°C). In some embodiments, the block face is maintained at approximately 10-12°C. In some embodiments, the humid air is at approximately room temperature. In some embodiments, the humid air is above approximately room temperature. In some embodiments, the humid air is below approximately room temperature.

[0113] In some embodiments of the present disclosure, the tissue block is cooled to maintain a temperature at the block face below about 12° C., as discussed above. In some embodiments of the present disclosure, the tissue block is cooled to below 10° C. in the microtome. Various embodiments of cooling methods and systems are disclosed above.

[0114] In some embodiments, in step 816, the hydration method includes spraying hydrating liquid onto the block face, depositing liquid hydrating liquid droplets on the surface of the block face. In some embodiments, a mechanical or pneumatic pump using a sprayer nozzle at a temperature below about 10° C. may be employed to spray the sample block. In some embodiments, the amount sprayed is generally sufficient to cover the block face with surface hydrating liquid at about room temperature for about a few seconds. The spraying presupposes a dedicated chamber that can contain many blocks, but this may not necessarily be equipped as a hydration chamber, at least for the liquid hydrating liquid spraying. Conversely, some directed source of moisture may be embodied within the hydration chamber using a more focused or intentionally directed spray of moisture onto the block face from an external source, i.e., a sonicator, or otherwise internal to the hydration chamber.

[0115] In some embodiments, a chemical additive can be applied to the block face to further facilitate or accelerate the process of aqueous diffusion into the tissue / wax composite medium. The block face may then be subjected to any form of humidification, including directed hydrating liquid droplet spray, focused streams of moisture (hydrating liquid vapor), or any other exemplary mechanical method that can serve as a source of hydrating liquid, in a form that includes both a liquid phase (hydrating liquid droplets) or a gas phase (water that tends to condense on the cold block face). Consistent with the specific (thermodynamic) activity of the chemical additive, aqueous diffusion is thereby accelerated, and the enhanced rate of tissue hydration may result from any number of mechanisms involving various physical or chemical forces, such as surface tension, diffusion, and wetting, that allow the hydrating liquid to overcome the transport resistance associated with the hydrophobic properties of the wax, i.e., its high contact angle with the hydrating liquid.

[0116] In some embodiments, an optional surfactant may be used in step 818. In step 820, an oil-soluble surfactant may be applied (e.g., brushed, sprayed) onto the block face in the hydration chamber so that a hydrating liquid-oil emulsion forms on the block face when water is applied to the block face. The surfactant can be applied directly to the block face to induce hydrating liquid penetration into the tissue / wax composite medium, resulting from W / O emulsification at the interface of rapidly condensing hydrating liquid droplets from moist air (e.g., at room temperature) on the extremely cold block face, maintained below the dew point of the condensing gas phase. In some embodiments, the block is operatively cooled in the hydration chamber by any number of exemplary means, including, but not limited to, the use of ice, a piezoelectric cold plate, dry ice, liquid nitrogen, etc. In some embodiments, a cooling system such as that described above is used. In this way, there is stabilization of the condensed droplets against coalescence so that the hydrated liquid droplets retain the integrity of their dispersed phase, i.e., they remain as dispersed hydrated liquid droplets upon merging with the continuous phase defined by the tissue / wax complex. While not wishing to be bound by theory, the mechanism is consistent with surfactants being generally more oil-soluble, which may allow them to more spontaneously form a homogeneous mixture with the paraffin wax, particularly at the air / block face interface, to the extent that several 4 micron slices per (hydrated) tissue block or tens of microns may be required to achieve sectioning of the tissue sample. A variety of oil-soluble surfactants can be used, including but not limited to sorbitan esters, glycerol stearate, polyethylene glycol oleyl ether, sorbitan monooleate, and sorbitan tristearate, or more generally, any oil-soluble surfactant that has a suitably low HLB value, as generally defined or generally understood to indicate the surfactant solubility in oily or hydrophobic medium versus that in hydrophilic medium (for example, less than about 8 HLB).In some embodiments, the concentration of surfactant is about 1 mM.

[0117] In some embodiments, the oil-soluble surfactant may be further dissolved in any oil-like substance, such as paraffin oil or wax. This two-component low-viscosity mixture may be applied to the exposed tissue of the block face, forcing the surfactant-laden oil so that the mixture infiltrates the tissue / wax medium, penetrates the embedded wax, and dissipates or diffuses into the wax at the exposed tissue / wax surface. In some embodiments, the surfactant is immersed in the wax. Paraffin oil, which matches the low MV end of the MW distribution for paraffin wax, is ideally compatible with higher MV waxes, and therefore simply serves to transport the surfactant, even if only to a depth of 50 microns. In some embodiments, a portion of the W / O surfactant (e.g., 10%) is mixed with pure paraffin oil and applied to the block face to entrain the W / O surfactant in the wax. In some embodiments, the surfactant may be entrained in the wax to a depth of approximately 20 microns. Furthermore, the hydration liquid vapor transmission rate is two orders of magnitude higher in liquid paraffin, thus facilitating the flow of hydration liquid across the interfacial boundary.

[0118] In some embodiments, accelerated hydration of tissue involves the use of one or more chemicals that are soluble in the hydration liquid, which can enhance the spreading and penetration of aqueous solutions, for example, by either liquid droplets in the form of a spray onto a block face or droplet condensation from a humidified chamber. These wetting agents are selected to reduce the surface tension of the hydration liquid to approximately 20-22 mN / m at very low application levels (e.g., less than about 2%), thereby allowing the aqueous solution to spread rapidly over the hydrophobic wax / tissue surface, a phenomenon known as superspreading. In some embodiments, surfactants are selected to result in a spreading exponent greater than about 0.5.

[0119] In some embodiments, even though embedded tissue has a higher tendency to absorb hydration liquid compared to pure wax, microchannels embodied in the (quenched) crystallized wax itself, which are typically prevented from being penetrated by hydration liquid due to their high surface energy, are now more likely to suffer from hydration liquid contamination.

[0120] A variety of super-spreading surfactants can be used in step 822. In some embodiments, such surfactants can include, but are not limited to, trisiloxane surfactants, which can consist of a siloxane chemical group and a hydrophilic alkyl ether tail, which can vary in length or composition.

[0121] Siloxane structure can vary in one form or another.For example, changing methyl end groups to hydroxyl can induce ballistic penetration of hydration liquid into tissue, and any ethoxylated siloxane surfactant is incorporated herein by reference as a representative chemical that is soluble in hydration liquid at very low use levels, allowing aqueous mixtures to exhibit the properties of superdiffusion, or more generally, enhanced penetration, into wax / tissue media utilized in histology for the preparation and fixation of biological tissues, for example, obtained as biopsies for use in histopathological analysis of tissue samples encountered in medical diagnosis.

[0122] The super-spreading surfactant can be selected from known and conventional super-spreading surfactants, such as organosilicon super-spreading surfactants disclosed in U.S. Pat. Nos. 7,507,775, 7,645,720, 7,652,072, 7,700,797, 7,879,916, and 7,935,842, and U.S. Patent Application Publication No. 2007 / 0131611 (the entire contents of which are incorporated herein by reference), and non-organosilicon super-spreading surfactants, such as those disclosed in U.S. Pat. Nos. 5,821,195, 6,130,186, 6,475,953, 7,723,265, and 7,964,552 (the entire contents of which are also incorporated herein by reference).

[0123] In some embodiments, a mixture of an organic silicone super-diffusing surfactant and a non-organic silicone super-diffusing surfactant can be employed as an optional component, as disclosed in the aforementioned U.S. Patent No. 7,964,552. Advantageously, additional surfactant combinations that can be included in the compositions of the present disclosure include an organic silicone super-diffusing surfactant and a non-super-diffusing organic silicone surfactant, an organic silicone super-diffusing surfactant and a non-super-diffusing non-organic silicone surfactant, a non-organic silicone super-diffusing surfactant and a non-super-diffusing organic silicone surfactant, a non-organic silicone super-diffusing surfactant and a non-super-diffusing non-organic silicone surfactant, and a non-super-diffusing organic silicone surfactant and a non-super-diffusing non-organic silicone surfactant. The weight ratio of different types of surfactants in these mixtures can vary widely, for example, from 1:100 to 100:1, preferably from 1:50 to 50:1, and more preferably from 1:20 to 20:1.

[0124] The organic silicon super-spreading surfactant can be selected from any one or more of the known and conventional tri- and tetrasiloxane alkoxylate types.Non-limiting examples of suitable surfactants are disclosed, for example, in U.S. Patent No. 9,034,960 (incorporated herein by reference in its entirety).Some polysiloxane alkoxylates (I), such as Silwet L-77 and Silwet 408, are commercially available from Momentive Performance Materials Inc.

[0125] The very low equilibrium surface tension of trisiloxane solutions is not the only reason for the superspreading behavior; the characteristic wetting properties of trisiloxane surfactants are superior, in part due to their better adsorption to hydrocarbon solid substrates, and therefore the solid / liquid interfacial tension is lower. Superspreading behavior can begin at a critical wetting concentration (CVC), which can be much higher than the critical coagulation concentration (CAC), and this behavior demonstrates the dynamic surface tension effect underlying the mechanism of superspreading. Note that the CWC is independent of the substrate used for spreading. In some embodiments, the surfactant concentration is less than about 2 wt.%.

[0126] Surfactant adsorption at air / liquid versus solid / liquid interfaces can be very different in terms of surfactant affinity for the surface because the chemical composition of each interface is uniquely different. In some embodiments, a single surfactant can be used. In some embodiments, instead of a binary mixture of a single surfactant in the hydrating liquid, any combination of surfactants may be used, where each surfactant may be categorically different in terms of chemical structure or some other defining characteristic. In this way, a synergistic effect may be achieved, providing better wetting properties.

[0127] Superspreading behavior generally correlates with lower surface tension, but that is not the only factor affecting overall performance. The size and structure of the actual surfactant molecule can also play an important role in spreading performance. Trisiloxane copolymers belong to specific classes according to the specific chemical structure of the surfactant molecule. Trisiloxane surfactants are block copolymers of silicone, ethylene oxide (EO), and / or propylene oxide (PO). Typically, they have either a pendant graft structure or a linear (ABA) structure. The specific composition depends on the individual magnitudes of the variables x, y, m, and n.

[0128] Molecules with significant silicone content will exhibit lubricity, desorption, low coefficient of friction, slipperiness, and abrasion resistance in various industrial or personal care applications. Molecules with high polyethylene oxide content will be self-dispersible or soluble in hydrating liquids or polar solvents. This reduces the interfacial tension, thus aiding wetting, spreading, and penetration. Materials with high polypropylene oxide content will be self-dispersible or soluble in non-polar solvents. This reduces the interfacial tension of such solvents, thus aiding wetting and flow.

[0129] In some embodiments, super-diffusing trisiloxanes with pendant graft structures can be used either alone or in combination with other surfactants.

[0130] In some embodiments, compositions are provided comprising formulations comprising one or more surfactants as described in U.S. Patent No. 8,734,821 (incorporated herein by reference in its entirety). The differentiation of adsorption kinetics at the air / liquid versus solid / liquid interface, or even the dynamic adsorption of surfactant molecules in front of the three-phase contact line at the solid / air interface, influences and promotes the wetting process. Surfactant redistribution in mixed systems affects the rate of diffusion; therefore, in addition to the dynamic surface tension phenomenon, the equilibrium values ​​of surface and interfacial tension play a role in superdiffusion and therefore affect the rate of aqueous penetration into hydrophobic media, such as wax / tissue composites used in histological analysis of biological tissue samples derived from surgical biopsies of both normal and diseased tissues to make medical diagnoses regarding the origin or nature of disease by pathologists and other medical professionals. In some embodiments, the dynamic surface tension is less than about 22 dynes / cm.

[0131] In some embodiments, the processing of tissue blocks in an automated device is improved by providing a spraying process therein, in which a super-diffusing mixture is applied directly to the block face to accelerate the transition from sectioning to hydration, thus not only improving the speed of processing, but also simplifying the overall mechanical design of the device insofar as the number of moving parts or distinct or discrete mechanical steps is significantly minimized.

[0132] 9-12 provide non-limiting examples of suitable hydration systems. Referring to FIG. 9, hydration system 900 may include a hydration chamber 901 in communication with a humidifier 902. Humidifier 902 is in fluid communication with hydration chamber 901 and supplies moist air 904 to the hydration chamber. Condensate 906 can be vented out of hydration chamber 901.

[0133] 10 , in some embodiments, the humidifier 902 can include a basin 918 for storing hydration liquid and a lid 914. In some embodiments, the humidifier 902 can be an ultrasonic humidifier for providing air with high humidity to the hydration chamber 901 via an ultrasonic disk 910 and a blower 912. Condensate can be returned to the humidifier via a condensate port 916.

[0134] Referring to FIG. 11 , the humidifier chamber 901 includes a housing 938 designed to receive one or more tissue blocks 930. The moist air 904 can condense on the surfaces of the tissue blocks 930, as discussed above. Unabsorbed condensation can be returned to the humidifier 902 through additional tubing. In some embodiments, the hydration chamber 900 can include a retractable lid 932, which allows removal of the tissue block 930 from the hydration chamber and / or can adjust the humidity within the hydration chamber. In some embodiments, a seal motor 934 can be provided to seal the hydration chamber while the tissue block is hydrated. In some embodiments, the hydration chamber 901 also includes a drive gear 936 for moving the tissue block through the hydration chamber. Because the interior of the hydration chamber 901 is initially dry, overflowing condensation can be collected and reused.

[0135] 12 , in some embodiments, the tissue block can be hydrated in one or more microtomes. In such embodiments, the system may include a humidifier 1202, similar to humidifier 902, for example. The humidifier can be positioned proximate to the microtome chuck 1203 and can use a hose or nozzle 1204 to provide hydrating liquid directly onto the block face of the tissue block 1230 positioned within the microtome chuck 1203. Unabsorbed condensation can be discarded.

[0136] Any suitable computing device may be used to implement the computing devices and methods / functionality described herein and, as will be understood by those skilled in the art, may be converted into a specific system for performing the operations and features described herein through hardware, software, and firmware modifications in a manner that goes significantly beyond simply running software on a typical computing device. One illustrative example of such a computing device 1300 is depicted in FIG. 13. Computing device 1300 is merely an illustrative example of a suitable computing environment and does not limit the scope of the present disclosure in any way. A "computing device" as represented by FIG. 13 may include a "workstation," a "server," a "laptop," a "desktop," a "handheld device," a "mobile device," a "tablet computer," or other computing device, as will be understood by those skilled in the art. Given that computing device 1300 is depicted for illustrative purposes, embodiments of the present disclosure may utilize any number of computing devices 1300 in any number of different ways to implement a single embodiment of the present disclosure. Thus, embodiments of the present disclosure are not limited to a single computing device 1300, or to a single type of implementation or configuration of an exemplary computing device 1300, as will be understood by one of ordinary skill in the art.

[0137] Computing device 1300 may include a bus 1310 that may be coupled, directly or indirectly, to one or more of the following illustrative components: memory 1312, one or more processors 1314, one or more presentation components 1316, input / output ports 1318, input / output components 1320, and power supply 1324. Those skilled in the art will understand that bus 1310 may include one or more buses, such as an address bus, a data bus, or any combination thereof. Those skilled in the art will additionally understand that, depending on the intended application and use of a particular embodiment, multiple of these components may be implemented by a single device. Similarly, in some cases, a single component may be implemented by multiple devices. Thus, FIG. 13 is merely an illustration of an exemplary computing device that may be used to implement one or more embodiments of the present disclosure and does not limit the disclosure in any way.

[0138] Computing device 1300 may include or interact with various computer-readable media. For example, computer-readable media may include random access memory (RAM), read-only memory (ROM), electronically erasable programmable read-only memory (EEPROM), flash memory or other memory technology, CD-ROM, digital versatile disk (DVD) or other optical or holographic media, magnetic cassettes, magnetic tape, magnetic disk storage, or other magnetic storage devices that may be used to encode information and that may be accessed by computing device 1300.

[0139] The memory 1312 may include computer storage media in the form of volatile and / or nonvolatile memory. The memory 1312 may be removable, non-removable, or any combination thereof. Exemplary hardware devices are devices such as hard drives, solid-state memory, optical disk drives, and the like. The computing device 1300 may include one or more processors that read data from components such as the memory 1312, various I / O components 1316, and the like. The presentation component 1316 presents an indication of the data to a user or other device. Exemplary presentation components include a display device, a speaker, a printing component, a vibrating component, and the like.

[0140] I / O ports 1318 may allow computing device 1300 to be logically coupled to other devices, such as I / O components 1320, some of which may be built into computing device 1300. Examples of such I / O components 1320 include microphones, joysticks, recording devices, gamepads, satellite dishes, scanning devices, printers, wireless devices, networking devices, and the like.

[0141] The examples provided to aid in understanding the present disclosure should not be construed in any way as limiting the scope of the disclosure as defined in the claims that follow. The following examples are provided to provide those of ordinary skill in the art with a complete disclosure and description of how to make and use embodiments of the present disclosure, and are not intended to limit the scope of the disclosure or represent that the experiments described below are all or the only experiments performed. Efforts have been made to ensure accuracy with respect to numbers used (e.g., amounts, temperatures, etc.), but some experimental error and deviation should be accounted for.

[0142] Example

[0143] The trisiloxane surfactants employed in this example are Silwet L-77 and Silwet 408 (see, e.g., U.S. Pat. No. 8,734,821, incorporated herein by reference in its entirety). The formulations employed in this example were prepared according to the following procedure: The trisiloxane surfactant or surfactants, Silwet L-77 and / or Silwet 408, are added to purified water at 0.5% by weight. The aqueous mixture is easily blended. At this point, other ingredients, such as solvents or other additional surfactants, may be included in the formulation. Table 1 below presents the compositions of the formulations containing the indicated trisiloxane surfactants at equal concentrations, without any other ingredients being incorporated into the composition. [Table 1]

[0144] The following table presents the results of experiments monitoring block face hydration time in response to treatment with formulations prepared as described above employing the indicated trisiloxane surfactant or surfactants at equal concentrations. [Table 2]

[0145] As illustrated by the tabulated test results, it is clear that the reduction in hydration time achieved by employing the Silwet L-77 / Silwet 408 blend is a synergistic improvement compared to the results achieved by the individual component surfactants alone.

[0146] Numerous modifications and alternative embodiments of the present disclosure will be apparent to those skilled in the art in view of the foregoing description. Accordingly, this description is to be construed as illustrative only and is for the purpose of teaching those skilled in the art the best mode of carrying out the present disclosure. Details of construction may vary substantially without departing from the spirit of the present disclosure, and the exclusive use of all modifications that fall within the scope of the appended claims is reserved. While embodiments have been described herein in a manner that permits a clear and concise specification to be written, it is intended and should be understood that the embodiments can be combined or separated in various ways without departing from the scope of the present disclosure. It is intended that the present disclosure be limited only to the extent required by the appended claims and applicable legal provisions.

[0147] As used herein, the terms "comprises" and "comprising" are intended to be interpreted as inclusive, not exclusive. As used herein, the terms "exemplary," "example," and "illustrative" are intended to mean "serving as an example, instance, or illustration" and should not be interpreted as indicating a preferred or advantageous configuration over other configurations. As used herein, the terms "about," "generally," and "approximately" are intended to cover variations that may exist at the upper and lower limits of a range of subjective or objective values, such as variations in properties, parameters, sizes, and dimensions. In one non-limiting example, the terms "about," "generally," and "approximately" mean 10 percent or +10 percent or less, or -10 percent or less. In one non-limiting example, the terms "about," "generally," and "approximately" mean close enough to be considered included by one of ordinary skill in the art. As used herein, the term "substantially" refers to a complete or nearly complete extension or degree of an action, characteristic, property, state, structure, item, or result, as would be understood by one of ordinary skill in the art. For example, an object that is "substantially" circular would mean that the object is either perfectly circular to the limits of mathematical determination, or approximately circular as would be recognized or understood by one of ordinary skill in the art. The precise degree of acceptable deviation from absolute perfection may, in some cases, depend on the specific context. However, in general, near perfection would be such that one has the same overall result as if absolute and total perfection were achieved or obtained. The use of "substantially" is equally applicable when utilized in a negative sense to refer to a complete or nearly complete lack of an action, characteristic, property, state, structure, item, or result, as would be understood by one of ordinary skill in the art.

[0148] Numerous modifications and alternative embodiments of the present disclosure will be apparent to those skilled in the art in view of the foregoing description. Accordingly, this description is to be construed as illustrative only and is for the purpose of teaching those skilled in the art the best mode of carrying out the present disclosure. Details of construction may vary substantially without departing from the spirit of the present disclosure, and the exclusive use of all modifications that fall within the scope of the appended claims is reserved. While embodiments have been described herein in a manner that permits a clear and concise specification to be written, it is intended and should be understood that the embodiments can be combined or separated in various ways without departing from the present disclosure. It is intended that the present disclosure be limited only to the extent required by the appended claims and applicable legal provisions.

[0149] It is also to be understood that the following claims are intended to cover all general and specific features of the disclosure described herein, as well as all language within the scope of the disclosure that may be considered to fall therebetween as a matter of language.

Claims

1. 1. An automated system for preparing a tissue sample, the system comprising: one or more microtomes; A hydration system; A processor, the processor comprising: commencing sectioning of a first tissue block with one or more microtomes, the first tissue sample being embedded within an embedding material; hydrating the first tissue block with the hydration system for a first predetermined time period; commencing sectioning of a second tissue block with the one or more microtomes while the first tissue block is hydrated, the second tissue block comprising a second tissue sample embedded within an embedding material; hydrating the second tissue block with the hydration system for a second predetermined time period; starting the one or more microtomes to begin sectioning the first tissue block while the second tissue block is hydrated; a processor programmed to A system comprising:

2. The automated system of claim 1 , wherein the one or more microtomes comprise a first microtome and a second microtome.

3. The processor: commencing sectioning of the first tissue block with a first microtome; hydrating the first tissue block with the hydration system; commencing sectioning of the second tissue block by the first microtome while the first tissue block is in the hydration system; hydrating the second tissue block with the hydration system; commencing sectioning of the hydrated first tissue block with a second microtome while the second tissue block is hydrated; 10. The automated system of claim 1, programmed to:

4. The processor: commencing sectioning of the first tissue block with a first microtome; hydrating the first tissue block with the hydration system; commencing sectioning of the hydrated first tissue block with the first microtome; sectioning the second tissue block with a second microtome; hydrating the second tissue block with the hydration system; commencing sectioning of the hydrated second tissue block with the second microtome; It is programmed to 2. The automated system of claim 1, wherein at least one of the sectioning, hydration, and sectioning by the second microtome is performed in parallel with the sectioning, hydration, and sectioning by the first microtome.

5. 5. The automated system of claim 1, further comprising one or more transport medium units configured to transport one or more sections from the first tissue block and the second tissue block to one or more slides.

6. 5. The automated system of claim 1, further comprising one or more tissue block handlers configured to transfer the first tissue block and the second tissue block between the one or more microtomes and the hydration system.

7. 7. The automated system of claim 6, wherein the processor is programmed to communicate with the one or more tissue block handlers and control movement of the first tissue block and the second tissue block between the one or more microtomes and the hydration system.

8. The automated system of claim 1 , further comprising a tissue block tray configured to receive the first tissue block and the second tissue block.

9. The automation system of any one of claims 1 to 4, further comprising a cooling system.

10. The automated system of claim 1 , wherein the hydration system comprises a hydration chamber configured to receive the first tissue block and the second tissue block inside the hydration chamber.

11. 11. The automated system of claim 10, wherein the processor is further programmed to move the first tissue block and the second tissue block through the hydration system for the first and second predetermined times, respectively.

12. The automated system of any one of claims 1-4, wherein the hydration system is positioned proximate to the one or more microtomes and configured to hydrate the first tissue block and the second tissue block in the one or more microtomes.

13. 1. A method for batch processing tissue blocks, the method comprising: initiating, by a processor, sectioning of a first tissue block comprising a first tissue sample embedded within an embedding material by one or more microtomes controlled by said processor; hydrating, by the processor, the first tissue block for a first predetermined period of time; initiating, by the processor, sectioning with the one or more microtomes a second tissue block while the first tissue block is hydrated, the second tissue block comprising a second tissue sample embedded within an embedding material; hydrating, by the processor, the second tissue block for a second predetermined period of time; starting, by the processor, the one or more microtomes to begin sectioning the first tissue block while the second tissue block is hydrated; A method comprising:

14. 14. The method of claim 13, further comprising transferring, by a tissue block handler in communication with the processor, the first tissue block and the second tissue block from the one or more microtomes to a hydration system for hydration.

15. The method of claim 13 , wherein the one or more microtomes comprise a plurality of microtomes.

16. 14. The method of claim 13, further comprising transporting one or more sections sectioned from the first tissue block and the second tissue block from the one or more microtomes to one or more slides by one or more transport medium units in communication with the processor.

17. 1. A method for processing a tissue sample, the method comprising: sectioning, with a first microtome, a first tissue block comprising a first tissue sample embedded within an embedding material; hydrating the first tissue block with a hydration system for a predetermined period of time; sectioning a second tissue block with the first microtome while the first tissue block is hydrated, the second tissue block comprising a second tissue sample embedded in an embedding material; hydrating the second tissue block with the hydration system; sectioning a sample from the hydrated first tissue block with a second microtome while the second tissue block is hydrated; A method comprising:

18. 18. The method of claim 17, further comprising transferring one or more sections sectioned from the first tissue block from the second microtome to one or more slides.

19. 1. A method for processing a tissue sample, the method comprising: sectioning, with a first microtome, a first tissue block comprising a first tissue sample embedded within an embedding material; hydrating the first tissue block with a hydration system for a first predetermined period of time; sectioning the hydrated first tissue block with the first microtome; sectioning, with a second microtome, a second tissue block comprising a second tissue sample embedded within an embedding material; hydrating the second tissue block with the hydration system for a second predetermined period of time; sectioning the hydrated second tissue block with the second microtome; Including, A method wherein at least one of the sectioning, hydrating, and sectioning with the second microtome is performed in parallel with the sectioning, hydrating, and sectioning with the first microtome.

20. 20. The method of claim 19, further comprising transferring one or more sections sectioned from the first tissue block from the first microtome to one or more slides.

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