System and method for biological specimen mounting

A system and method for mounting a section onto a substrate, the system comprising: a fluid channel including: a fluid channel inlet that receives the section, processed from a bulk embedded sample by a sample sectioning module positioned proximal the fluid channel inlet, a section-mounting region downstream of the fluid channel inlet, and a fluid channel outlet downstream of the section-mounting region; a reservoir in fluid communication with the fluid channel outlet; and a manifold, fluidly coupled to the reservoir, that delivers fluid from the reservoir to the fluid channel inlet, thereby transmitting fluid flow that drives delivery of the section from the fluid channel inlet toward the section-mounting region.

TECHNICAL FIELD

This invention relates generally to the biological research field, and more specifically to a new system and method biological specimen mounting.

BACKGROUND

It is commonly desirable in biological laboratories to mount tissue sections, or ‘specimens’, to slides for purposes of examining the tissue sections using a microscope, treating the tissue sections with a stain or dye, and for other purposes. Conventional systems and methods for mounting specimens onto slides comprise placing tissue sections in a sufficiently deep water bath, with the specimens floating on the surface of the water. The broad side of a slide is then rested on the rim of the water bath and the slide is angled down into the water back such that the slide is partially submersed in the water. Subsequently, a small brush or glass capillary tube is used to manipulate a tissue section onto the slide. Typically, the slide is gradually drawn out of the water as additional tissue sections are arranged on the slide. In another variation of a conventional method, tissue is embedded in paraffin wax, sliced with a microtome, and then selected sections of the embedded tissue are transferred to a heated water bath. The hot water bath partially melts the paraffin about the specimens, and a glass slide treated with adherents is then used to scoop the tissue sections out of the hot water bath. Conventional methods of mounting specimens on slides are thus difficult, time-consuming, and labor-intensive.

There is thus a need in the biological research field for a new system and method for biological specimen mounting. This invention provides such a new system and method.

BRIEF SUMMARY OF THE INVENTION

Not Applicable.

DESCRIPTION OF THE EMBODIMENTS

The following description of preferred embodiments of the invention is not intended to limit the invention to these embodiments, but rather to enable any person skilled in the art to make and use this invention.

As shown inFIG. 1, an embodiment of a system100for coupling a section101to a substrate102comprises: a fluid channel110having a fluid channel inlet120that receives the section101, processed from a bulk embedded sample by a sample sectioning module103positioned proximal the fluid channel inlet120, a section-mounting region130downstream of the fluid channel inlet, and a fluid channel outlet140downstream of the section-mounting region; a reservoir150in fluid communication with the fluid channel outlet; and a manifold160fluidly coupled to the reservoir, that delivers fluid from the reservoir to the fluid channel inlet, thereby transmitting fluid flow that drives delivery of the section from the fluid channel inlet toward the section-mounting reservoir. In some embodiments, the system100can additionally or alternatively include any one or more of: a filter170, fluidly configured between the fluid channel outlet and the manifold, that prevents undesired substances from flowing into the fluid channel inlet; a temperature regulating module180in contact with fluid from the reservoir, that adjusts a temperature of fluid within the fluid channel; and a substrate actuation module190that transmits the substrate into the section-mounting region in a first operation, and delivers the substrate from the section-mounting region, with the section mounted to the substrate, in a second operation.

The system100functions to automate processing of sections (e.g., histological specimen sections, biological sections, etc.) in a manner that consistently generates high-quality mounted sections, with minimal or no effort from a human technician. As such, the system100can significantly reduce labor-intensive aspects of mounting sections to substrates. The system100is preferably configured to implement at least a portion of the method200described in Section 2 below.

In one specific workflow, the system100is configured to retrieve a thin tissue section (e.g., generated from a microtome blade), to separate the tissue section from a preceding section, to transport the section to a microscope slide via a fluidic channel, and then to mount the section onto the microscope slide with a substrate actuation module that coordinates movement of the microscope slide in relation to motion of the tissue section within the fluidic channel. In mounting a tissue section onto the microscope slide, the geometry of the fluidic channel is configured to deliver the tissue section toward an interface at which the microscope slide and the surface of fluid within the fluidic channel intersect, center the tissue section onto the microscope slide, and orient the tissue section such that its sides are parallel to long edges of the microscope slide. Mounting, in the specific workflow, is then consummated by causing a line of juncture between the microscope slide and the surface of the fluid within the fluidic channel to recede in a direction opposite to that of flow within the fluid section. In variations of the specific workflow, recession of the line of juncture to facilitate mounting can be accomplished by slowing flow of fluid (e.g., by decreasing a volumetric flow rate of fluid) within the fluidic channel, by providing relative motion between the fluidic channel and the microscope slide in a manner that enhances mounting of the tissue section to the microscope slide, by removing a previously-submerged displacing body from a fluid volume within the fluidic channel (i.e., to lower the fluid level within the fluid channel), and/or by any other suitable mechanism. In the specific workflow, the substrate actuation module can further be configured to modulate motion of the microscope slide to be positioned for placement of multiple sections onto the slide or to be fully retracted to create an unobstructed path to carry discarded sections to a reservoir for filtration and/or recirculation. The system100can, however, facilitate any other suitable workflow or method involving any other suitable section and/or imaging substrate.

In variations wherein the system100interacts or integrates with a sample sectioning module103, the system100can be configured to cooperate with the sample sectioning module103in order to separate serially connected sections generated by the sample sectioning module103for transmission into the fluid channel110. In one example of a sample sectioning module103comprising a microtome104, as shown inFIG. 2, a blade3(e.g., microtome blade) of the microtome104is retained in position by a blade holder having a stage22that collects tissue sections during normal operation. The microtome104can have an adjustable blade angle and an adjustable stage angle ox, as shown inFIG. 2, that coordinates with an angle of the blade. The stage22of the microtome104can thus rotate with an axis of rotation about the tip of the blade3, and the system100can mate with the stage22along an interface (e.g., linear interface) between the blade3and the system100such that the blade angle can be adjusted without repositioning of the system100. Furthermore, this configuration allows for lateral adjustment of the blade3within the microtome, without repositioning of the system100in relation to the microtome104. The system100can further be hermetically sealed against the stage22at the fluid channel110or manifold160(e.g., using a sealing gasket, using mechanical pressure, etc.) in order to minimize fluid leakage at an interface between the stage22and the fluidic channel110. In one alternative to the specific example, the system100can directly interface with the stage22or another portion of the blade-holding portions of the microtome104. In another alternative to the specific example, the system100can include portions that substitute for the stage22and couple directly to blade-holding portions of the microtome104. The sample sectioning module103can, however, include any other suitable elements or be configured relative to the system100in any other suitable manner.

In the example above, each cut motion of the microtome104produces a new section101, and the embedding material used for the section101preferably has a density lower than that of fluid (e.g., water) flowing through the system100, such that the section100floats on the surface of the fluid. Preferably, each generated section101remains coupled to the blade3(e.g., loosely coupled to the blade by way of the embedding medium), and fluid introduced through a manifold160into the fluid channel110at an angle γ frees a preceding section for transmission through the fluid channel110and mounting. Flow at the angle γ frees the preceding section by providing a force that produces tension at a junction between serial sections generated at the microtome104. Additionally, in a related example, a portion of fluid flow from the manifold160is directed to flow against the stage22and in a superior direction towards the blade3, which facilitates uniform pulling of sections away from the blade3as they are cut by the blade3. Furthermore, in the related example, features (i.e., fins) oriented with a direction of fluid flow within the fluid channel110at the fluid channel inlet120promote laminar flow away from the blade3.

Additionally or alternatively, separation of a section101from the blade3can be performed by generating fluid flow beneath a section101within the fluid channel110, such that a shear force induced at a junction between sections provides separation. Still alternatively, an operator can manually separate a section101from the blade3(e.g., using forceps). Still alternatively, an elevated floor of the fluid channel inlet120, immediately downstream of the manifold160, can cause fluid to be drawn away from the blade3as it is delivered into the fluid channel100. Such a configuration, as shown inFIGS. 3A and 3B, enables a cushion of water to develop near the blade3with high flow rates, and can allow multiple sections to be separated using flow speed modulations that retain a section attached to the blade3, while biasing a preceding section away from the blade3. Still alternatively, as shown inFIG. 3C, a concave surface111of the fluid channel inlet120can provide a “bowl” of fluid that facilitates retention of a section attached to the microtome blade3, while openings of the manifold160project fluid underneath the section to facilitate separation of adjoining sections. Multiple orifice angles, as shown inFIG. 3C, can provide a force that facilitates flexing of adjoining sections, thereby promoting separation from a shear force induced at a junction between adjoining sections.

Still alternatively, a separation device of the system100(e.g., a paddle, a chuck, a solenoid plunger, etc.) can use mechanical force to separate adjoined sections. In one example, as shown inFIG. 4A, fluid flow can modulate motion of a separation device72ain separating adjoined sections and allowing a released section to be transmitted into downstream portions of the fluidic channel110. In another example with a paddle72b, as shown inFIG. 4B, as the microtome chuck rises, a band71connecting a lever arm on the paddle72bto the chuck can pass above the paddle's pivot point, causing the paddle to transition to an active configuration. Then, the paddle72bcan be configured to revert to an inactive configuration, as shown inFIG. 4C, when the chuck descends as sections are being sliced from the bulk embedded sample. In yet another example, a solenoid plunger configured proximal the fluid channel inlet120can provide a force that separates a section from an adjoining section.

Once a section101has been separated in any of the above variations and examples, a shallower depth25within the fluid channel110, as described in further detail below, can allow the section101to accelerate toward downstream portions of the fluid channel110. In any of the above examples, having a section101adhere to the blade3for a brief period of time prior to separation by fluid transmission allows an operator to observe its quality and intervene in the sample processing process, if necessary. Floating a section101atop fluid in the fluid channel110, with coupling of the section101to the blade3can further function to reduce the presence of any wrinkling in the section101. Additionally or alternatively, in any of the examples,

The fluid channel110has a fluid channel inlet120that receives the section101, processed from a bulk embedded sample by a sample sectioning module103positioned proximal the fluid channel inlet120, a section-mounting region130downstream of the fluid channel inlet, and a fluid channel outlet140downstream of the section-mounting region. The fluid channel110functions to receive the section101from a sample sectioning module103, and to deliver the section over a layer of flowing fluid that drives the section for mounting at a downstream position. The fluid channel110preferably defines a primarily straight flow path; however, in some variations, the fluid channel110can alternatively define a curved flow path or any other suitable flow path. Preferably, the fluid channel110is wider than a maximum width of the section in order to facilitate smooth transmission of the section into the fluid channel110(e.g., to prevent jamming) during delivery along the fluid channel110. However, the fluid channel can alternatively have any other suitable width relative to a width of the section. Furthermore, the width and/or depth of the fluid channel110can be constant or variable, in order to produce desired flow behavior through portions of the fluid channel110. As such, constricted portions of the fluid channel110can produce higher velocities of fluid flow than less constricted portions of the fluid channel110, given a volumetric flow rate of fluid through the fluid channel110. In some variations, the fluid channel110can have at least one declined portion relative to a horizontal plane in order to passively facilitate fluid flow. In some variations, the fluid channel110can additionally or alternatively comprise portions that are flat or inclined relative to a horizontal plane.

The fluid channel inlet120is preferably configured proximal to an output region of the sample sectioning module103, in order to facilitate initial positioning of the section, from the sample sectioning module103, within the fluid channel inlet120. In specific examples, as shown inFIGS. 2 and 5, the fluid channel inlet120is configured proximal to a blade3(e.g., a stationary blade) of a microtome, wherein interaction between a bulk embedded sample (i.e., a biological sample embedded in wax) and the blade generates the section and delivers the section toward the fluid channel inlet120. In the specific example, the bulk embedded sample is configured to couple to an actuator that moves the bulk embedded sample relative to the stationary blade to generate sections; however, variations of the specific example can involve any other suitable relative motion between a bulk embedded sample and a cutting instrument to generate sections. As such, the system100can be configured to couple directly to or to be positioned adjacent to an output region of a sample sectioning module103; however, the system100can additionally or alternatively be configured such that a user or other entity can transfer a section generated from any suitable sectioning device to the fluid channel inlet120for histological mounting.

The fluid channel inlet120preferably has a width substantially larger than that of a section101generated from a bulk embedded sample, in order to prevent wrinkling or any other form of damage to the section101upon transmission into the fluid channel inlet120. In variations, the fluid channel inlet120can have a width that is from 115% to 300% of the width of a section101generated by the sample sectioning module103. However, the width of the fluid channel inlet120can alternatively be any other suitable size in relation to a width of a sample generated at the sample sectioning module103. Furthermore, the width of the fluid channel110can be modulated from the fluid channel inlet120, to the section-mounting region130, to the fluid channel outlet140, in order to facilitate focusing and/or accurate positioning of a section101onto a substrate102at the section-mounting region130; however, the width of the fluid channel110can alternatively be substantially constant across two or more of the fluid channel inlet120, the section-mounting region130, and the fluid channel outlet140.

In some variations, the fluid channel inlet120can comprise a junction125at an upstream portion of the fluid channel inlet120, as shown inFIG. 6A, such that the junction125diverts a direction of fluid flow into the fluid channel100. As such, the junction125can function to provide a more compact and non-interfering interface between the fluid channel110and the sample sectioning module103. In one example, the junction is a 90° junction that allows a section transmitted into the fluid channel110to be diverted by an angle of approximately 90° between the sample sectioning module103and the fluid channel inlet120. Such a configuration facilitates positioning of the system100to interface with the sample sectioning module103in a first configuration (e.g., a coupled configuration), and facilitates removal of the system100from interfacing with the sample sectioning module103in a second configuration (e.g., a decoupled configuration). However, in alternative variations of the example, an angle of rotation between the fluid channel inlet120and the sample sectioning module103, provided by the junction125and defined inFIG. 6Aas θ, can alternatively range from 45° to 315°, or can have any other suitable angle depending upon morphological parameters of the fluid channel110and/or the sample sectioning module103.

In some variations, the junction125can define a region with a raised floor126, in relation to a manifold160, as described in further detail below. The raised floor126functions to provide concentration of fluid flow into the fluid channel inlet120, which allows acceleration of a section101floating atop and/or carried by fluid within the region of the junction125having a raised floor126. As such, the raised floor126can provide an inlet reservoir that provides desired initial motion characteristics (e.g., velocity, acceleration, flow path, etc.) of a section101entering the fluid channel110. Additionally or alternatively, the junction125can define a region that enables concentration of fluid flow into the fluid channel inlet120by defining a constricted cross-sectional area, perpendicular to a direction of fluid flow in the fluid channel inlet120, in any other suitable manner. For instance, a width and/or depth of a region of the junction125can be decreased within the junction125, relative to other portions of the fluidic channel110, thereby concentrating fluid flow into the fluid channel inlet120and accelerating motion of a section101within the junction125for a given volumetric flow rate in the junction. In related variations, a curved region of the junction125of the fluid channel inlet120(e.g., the raised floor region126) can include a set of tracks26, a specific example of which is shown inFIG. 6B, wherein the set of tracks divide the curved region of the junction125into a set of regions with varying fluid heights. The set of tracks26thus allow fluid travelling along the outside of the curved region of the junction125(e.g., fluid travelling the greatest distance) to move faster, thereby fluidically rotating a section101as it rounds the curved region of the junction125. This preserves an orientation of the section101(e.g., in relation to an orientation from the bulk embedded sample) and prevents jamming of sections within the system100.

In some variations, an output region of the fluid channel inlet120(e.g., defined at an output region of the junction125) can include a lip127(e.g., an elevated lip) protruding from a base surface of the fluid channel inlet120/junction125, that directs fluid, with a section101, into portions of the fluid channel110downstream of the fluid channel inlet120. The lip127can thus provide desired initial motion characteristics (e.g., velocity, acceleration, flow path, etc.) of a section101entering portions of fluid channel110downstream of the lip127, such that sections travelling within the fluid channel110travel in a predictable and/or repeatable manner. The fluid channel inlet120and/or junction125can, however, include any other suitable features that provide predictable flow behavior (e.g., substantially constant streamlines) that drives motion of sections within the fluid channel110.

The section-mounting region130of the fluid channel110is preferably a region of the fluid channel110configured between the fluid channel inlet120and the fluid channel outlet140, such that a section101transmitted into the fluid channel110by way of the sample sectioning module103is configured to be mounted to a substrate102at a region of the fluid channel110downstream of the fluid channel inlet120and upstream of the fluid channel outlet140. Preferably, the section-mounting region130has a depth that can accommodate passage of an imaging substrate under a section (e.g., by way of the substrate actuation module190) within the section-mounting region130, without disturbance (e.g., wrinkling, damage) of the section. In an example, as shown inFIG. 8, the section-mounting region130comprises a section-mounting reservoir132that is substantially deeper than the depth of the fluid channel inlet120and that allows a substrate to be submerged to a sufficient depth below a section101that has been delivered into the section-mounting region130. However, the section-mounting region130can alternatively be configured in any other suitable manner.

Preferably, the section-mounting region130is fluidly coupled to the fluid channel inlet120by a chute135, as shown inFIG. 6A, that functions to transport sections from the fluid channel inlet120to the section-mounting region130in a predictable and repeatable manner. The chute135also functions to provide desired motion characteristics (e.g., velocity, acceleration, flow path, etc.) of a section101upon delivery into the section-mounting region130, such that sequential sections travelling to the section-mounting region130reach the section-mounting region130in a consistent and desired manner. In one variation, the chute135can be oriented with a constant slope, defined inFIG. 6Aas P, that provides downhill flow for acceleration of a section101from the fluid channel inlet120to the section-mounting region130, as facilitated passively by gravitational force. Furthermore, in variations, the chute135can have an adjustable angle, such that the value of β can be adjusted (e.g., using an actuator coupled to the chute135or another portion of the fluidic channel110). In specific examples, β has a value from 5-15°, and in variations of the specific examples, β can have a value from 0-60° to provide desired flow characteristics within the chute135.

Alternatively, the chute135can have a varying slope along the length of the chute135, from an upstream portion to a downstream portion of the chute135, such that a profile of the chute135in an elevation view has a non-linear (e.g., curved) morphology. In one example, an upstream portion of the chute135has a steep slope (e.g., greater than 60°) relative to a horizontal plane, and the slope of the chute transitions to a substantially flat slope (e.g., less than 2°) in coupling to the section-mounting region130.

The chute135preferably facilitates focusing and accurate positioning of a section101at the section-mounting region, by having a width dimension that is reduced (e.g., gradually reduced) from the fluid channel inlet120to the section-mounting region130. In variations, the width of a downstream portion of the chute135, proximal the section-mounting region130, has a dimension that is from 105% to 125% of the width of a section101generated by the sample sectioning module103, such that the width of the downstream portion of the chute is substantially reduced relative to the width of the fluid channel inlet120. However, the width of the chute135can alternatively be any other suitable size in relation to a width of a sample generated at the sample sectioning module103. Additionally or alternatively, accurate positioning of a section traveling along the chute135can be facilitated by generating one or more well-defined streamlines of fluid flow, using channel morphologies that provide hydrodynamic focusing. In one example, the chute135can define a curved path that enables hydrodynamic focusing of a section101to a well-defined position at the section-mounting region130. In the example, the curved path can have a set of undulations that focus the section101from a not-well-defined position to a well-defined position in a consistent manner. Alternatively, a sonic steering module positioned at any portion of the fluid channel110can facilitate accurate positioning of a section. Still alternatively, accurate positioning of a section101at the section-mounting region130can be facilitated, by way of the chute135, in any other suitable manner.

In some variations, the section-mounting region130can include a base surface having a geometric feature131, as shown such that the geometric feature131is submerged below a fluid line of fluid within the fluid channel110, and provides flow characteristics that facilitate mounting of a section101to a substrate102at the section-mounting region130. In one such variation, the geometric feature131comprises a contoured surface133configured to align a section101passing over the geometric feature131, by way of fluid flow into the section-mounting region130, toward a desired position. In aligning the section101, the contoured surface produces a force vector that biases the section101against a substrate102within the section-mounting region130and aligns the section101such that its sides are substantially parallel with long edges of the substrate102. Alternatively, the section-mounting region130may omit a geometric feature131at a base surface, while still enabling mounting of a section101to a substrate102at the section-mounting region.

The fluid channel outlet140is preferably configured downstream of the section-mounting region130, in order to provide an outlet for flow from the fluid channel110. The fluid channel outlet140is preferably also configured to facilitate retrieval and/or filtration of undesired sections from the fluid channel110. As such, in some variations, the fluid channel outlet140can be configured to couple to a filtration and recirculation module that allows fluid and undesired sections from the fluid channel110to be filtered of the undesired elements, while allowing recirculation of fluid throughout the system (e.g., by way of the reservoir150). However, the fluid channel outlet140can alternatively be configured in any other suitable manner.

In one variation, an example of which is shown inFIGS. 6 and 8, the fluid channel outlet140comprises a curved spout142that allows fluid passing through the section-mounting region130to pass into a reservoir150that recirculates fluid back into the fluid channel110. Alternatively, the fluid channel outlet140can have any other suitable morphology that allows fluid from the section-mounting region130to pass through the fluid channel outlet140and into the reservoir150for recirculation. For instance, the fluid channel outlet140can include one or more of: a non-curved spout, a funnel-shaped feature, a manifold, and any other suitable fluid guiding feature that allows fluid to be efficiently delivered into the reservoir150(e.g., without leakage, without loss). The fluid channel outlet140is preferably configured to receive fluid passing through the section-mounting region130and to deliver fluid into the reservoir150whether or not a substrate102is present within the section-mounting region120; however, the fluid channel outlet140can alternatively be substantially obstructed when a substrate102is present within the section-mounting region130. The fluid channel outlet140is preferably elevated relative to the reservoir150, such that fluid from the fluid channel outlet140is passively delivered into the reservoir150as facilitated by gravity; however, the fluid channel outlet140can alternatively be configured relative to the reservoir in any other suitable orientation, wherein a driving element (e.g., pump) facilitates fluid flow from the fluid channel outlet140and into the reservoir150.

In a specific example of the fluid channel110′, as shown inFIGS. 6B and 7, the fluid channel inlet120′ includes a junction125′ having a region with a raised floor126′, in relation to a manifold160, that enables acceleration of a section101floating atop and/or carried by fluid within the fluid channel inlet120′. In the specific example, a curved region of the junction125, at the raised floor region126, includes a set of tracks26, wherein the set of tracks divide the curved region of the junction125into a set of regions with varying fluid heights, as shown inFIG. 6B. The set of tracks26in the specific example allow fluid travelling along the outside of the curved region of the junction125(e.g., fluid travelling the greatest distance) to move faster, thereby fluidically rotating a section101as it rounds the curved region of the junction125. In the specific example, the fluid channel inlet120′ also includes an elevated lip127′ protruding from a base surface of the fluid channel inlet125′, that directs fluid, with a section101, into portions of the fluid channel110downstream of the fluid channel inlet120. In the specific example, the fluid channel110is substantially straight between the output region of the fluid channel inlet120′ and the fluid channel outlet140, but rotated by 90° at the junction125′, in order to provide a more compact and non-interfering interface with the sample sectioning module103.

In the specific example of the fluid channel110′, the fluid channel110includes a chute135′ fluidly coupled between the fluid channel inlet120′ and the section-mounting region130′, wherein the chute135′ is configured to slope in a declined manner from the elevated lip127′ of the fluid channel inlet120; toward the section-mounting region130′. As such, the chute135′ provides downhill flow for acceleration of the section with fluid in the fluid channel110′. The slope of the declined portion is defined inFIG. 2as β and is defined as being from 5-15° in the specific example, and the section-mounting region130′ is substantially flat relative to a horizontal plane, such that the slope β of the fluid channel110transitions from being declined upstream of the section-mounting region130′ to being flat, relative to a horizontal plane, at the section-mounting region130′.

In the specific example of the fluid channel110, the channel width is initially substantially wider than (e.g., 115-300%) the width of a section101generated at the sample sectioning module103, but this width is then reduced, proximal to the section-mounting region130′, to a width that is marginally wider (e.g., 105-125%) than the width of the section101by sidewall contours of the fluid channel, in order to enable more accurate positioning of the section within the section-mounting region130. In the specific example of the fluid channel, the section-mounting region130comprises a receiving area including a contoured surface133at a base surface of the receiving area that is configured provide a biasing force that aligns the section toward a desired position at a substrate102within the section-mounting region130. Variations of the specific example of the fluid channel110can, however, be configured in any other suitable manner and comprise any other suitable fluidic elements that enable accurate and repeatable positioning of sections at one or more substrates within the section-mounting region130. Adjustable sidewall profiles, for instance, and as shown inFIG. 10, can be used to alter an amount of flow restriction about a substrate102to control fluid level heights, control fluid level modulation rates, accommodate samples of varying size, and/or adjust lateral positioning of a section101at a substrate102.

As noted above, the reservoir150is in fluid communication with the fluid channel outlet140, and functions to provide a bath of fluid that can be delivered into the fluid channel110by way of the fluid channel inlet120. The reservoir is preferably configured to receive fluid (e.g., filtered fluid) from the fluid channel outlet140for recirculation into the system, in order to enable reuse of a substantially fixed volume of fluid flowing throughout the system100. As such, the system100preferably includes a single reservoir that allows for fluid recirculation, wherein the single reservoir can be refilled, if needed (e.g., due to fluid loss in evaporation, etc.). However, variations of the system100can alternatively include any suitable number of reservoirs (e.g., a reservoir for fluid delivery into the fluid channel inlet120and a waste reservoir configured to receive waste fluid from the fluid channel outlet140) that enable fluid flow into the fluid channel inlet120and fluid flow out of the fluid channel outlet140.

The reservoir150preferably contains a volume of fluid that has desired properties in facilitating transmission of a section101along the fluid channel110, and mounting of the section101onto a substrate102at the section-mounting region130. In variations, the fluid can be characterized as one or more of: low-viscosity (e.g., less than 1×10−3Pa*s), volatile at temperatures for histological section processing (e.g., volatile at room temperature, volatile within a sample drying environment), non-interacting with histological process reagents (e.g., histological stains, etc.) to prevent generation of specimen artifacts, non-damaging to a biological specimen of a sample, neutral pH, and inexpensive. Preferably, the fluid circulating through the system100, by way of the reservoir150, comprises water; however, alternative variations of the fluid can comprise any other suitable fluid for histological section processing. In some variations, an additive can be introduced and/or neutralized to modify surface tension of the fluid to promote better transport of a section101through the fluidic channel, and/or to promote enhanced interactions with a substrate102. In one such example, a hydrophilic additive can be introduced with fluid from the reservoir to promote improved transport of a section101through the fluidic channel110.

The manifold160is fluidly coupled to the reservoir150, and functions to delivers fluid from the reservoir150to the fluid channel inlet120, thereby transmitting fluid flow that drives delivery of the section from the fluid channel inlet110toward the section-mounting region130. The manifold is configured to provide a flow path between the reservoir150and the fluid channel inlet120, thereby enabling separation of a section101from an adjoining section produced by the sample sectioning module103, facilitating delivery of the section101from the fluid channel inlet120, and transmitting the section toward the section-mounting region130of the fluid channel110. Preferably, fluid from the reservoir150is pumped through one or more tubes159into the manifold160, as shown inFIG. 6A, wherein the manifold160is configured to divide the flow into a set of openings162into the fluid channel inlet120. As such, the manifold160is preferably configured to generate laminar flow at the fluid channel inlet120; however, the manifold160can alternatively be configured to generate any other suitable type of flow (e.g., turbulent flow) at the fluid channel inlet120.

In one variation, as shown inFIG. 11, the manifold160has at least two inlet tubes159into the manifold160that provide a uniform (e.g., symmetric) distribution of flow across the openings162of the manifold160with negligible flow resistance. In this variation, the inlet tubes159are oriented in an opposing manner at opposite sides158of the manifold, wherein the opposite sides158are substantially parallel with sidewalls defining a longitudinal axis of fluid flow through the fluid channel110. In this variation, the set of openings162can be arranged in one or more of: a linear manner that defines a plane of fluid flow substantially parallel to that of a base surface of the fluid channel110; a linear manner that defines a plane of fluid flow substantially non-parallel to that of a base surface of the fluid channel110; a non-linear manner (e.g., curved manner defining a concave surface of fluid flow, curved manner defining a convex surface of fluid flow, staggered manner, etc.); and in any other suitable manner. In another variation, the manifold160can have an elongated opening163and/or an orifice pattern with a suitably apodized density, which can function to provide laminar flow into the fluid channel inlet120and distribute it more evenly across the opening(s)162, thereby eliminating the need for a second inlet tube159into the manifold. In another variation, the openings of the set of openings can be integrated into a single tube at varying angles in order to facilitate manipulation (e.g., separation) of the tissue sections. In yet another variation, as shown inFIG. 12, the manifold160can comprise a cavity164inferior to a surface with openings162, wherein fluid from the reservoir150is delivered into the cavity164, thereby allowing distribution of the set of openings162of the manifold across a surface (e.g., a planar surface parallel to a base surface of the fluid channel110) in a 2D or 3D configuration, rather than in a linear configuration. In yet another variation, the manifold160can be configured to deliver fluid from the reservoir150to one or both of the sidewalls of the fluid channel100, to produce flow in a direction non-parallel to a longitudinal axis of the fluid channel110. However, the manifold160can be configured to deliver fluid from the reservoir150and into the fluid channel inlet120using any other suitable 1D, 2D, or 3D configuration of openings162, or in any other suitable manner.

The manifold160is preferably in fluid communication with a pump167coupled between the reservoir150and the manifold, as shown inFIG. 1, wherein modulation of behavior of the pump167is governed by a controller168. The pump167can be configured to provide positive pressure and/or negative pressure in driving fluid between the reservoir150and the manifold160. As such, in one mode, forward flow generated by the pump167can facilitate forward movement of a section101through any portion of the fluid channel110, and in another mode, reverse flow generated by the pump167can facilitate reverse movement of a section101through any portion of the fluid channel110. Furthermore, forward and/or reverse flow can be adjustable to provide desired flow parameters (e.g., velocities, etc.) for processing of a single section or multiple sections in sequence. Modulation of flow (e.g., with a brief period of elevated flow rate) can additionally or alternatively be used to provide a biasing force that delivers a section101toward a substrate102for mounting.

In one variation, the pump167is a positive displacement pump, and in an example of this variation, the pump167is a peristaltic pump. In other examples, the pump167can include any one or more of: a gear pump, a screw pump, a piston pump, a progressing cavity pump, a roots-type pump, a plunger pump, a diaphragm pump, a rope pump, an impeller pump, and any other suitable type of pump. Furthermore, the system100can include more than one pump167configured at desired positions relative to the fluidic channel110, the reservoir150, and the manifold160. The pump167preferably has a known flow rate to pump speed ratio, such that control of the speed of the pump167corresponds to a control of the flow rate of the fluid within the fluid channel110. Furthermore, the pump167is preferably configured within the system100such that the system100is relatively easy to assemble, light to haul, quick to control, and easy to clean.

The controller168is preferably configured to respond to inputs provided by an operator of the system100, in modulating flow parameters of fluid within the system100. In one variation, the controller168can be configured to access a lookup table that facilitates correlation of an input from an operator of the system100to a desired flow parameter (e.g., flow rate) of the fluid within the fluid channel110. The lookup table preferably includes data based on one or more of: historical behavior of the system100, historical runs of other units of the system100, empirical data conducted and developed by the manufacturer or developer of the system100, and any other suitable data. The stored information preferably includes the type of fluid circulating throughout the system, characteristics (e.g., dimensions, embedding medium, etc.) of sections generated by a sample-sectioning module103in communication with the system100, number of sections being processed by the system100at a given time, potential errors in performance by the system100, and any other suitable information. The controller30can also be further adapted to access the lookup table via a computer processing network.

In another variation, the controller168can include a storage device with accessible memory. A user interface at which an operator provides inputs for control of the system100, along with the accessible memory of the storage device, can thus permit the operator to access stored information about runs of the system100and the system configuration and settings that were utilized during those runs. The stored information can include one or more of: the type of fluid circulating throughout the system, characteristics (e.g., dimensions, embedding medium, etc.) of sections generated by a sample-sectioning module103in communication with the system100, number of sections being processed by the system100at a given time, a history of errors in performance by the system100, and any other suitable information. This stored information can be accessed by the operator and retrieved by the controller168and/or systems. The operator can then, by interfacing with the controller168, automatically set up the flow parameters for the system100, by utilizing those previous sample run settings. Furthermore, once a run of the system100has been completed, an operator can save the controller settings and use the saved information for future runs for processing similar sections or specimens.

Flow modulation within the system100can, however, be additionally or alternatively enabled by using one or more valves that adjust flow (e.g., redirect flow, stop flow, open flow, etc.) to different reservoirs (e.g., a buffer reservoir) within or relative to the fluid channel100. Additionally or alternatively, a gate can be used to temporarily block passage of flow upstream of the section-mounting region130, thereby creating a desired drop in fluid level at the section-mounting region130, independent of a speed of operation of the pump167. The gate can also function to prevent a section from drifting back in an upstream direction. In operation, if pump speed remains unaltered, a fluid level on an upstream side of the gate will temporarily rise until the gate is removed from an obstructing position. Automation of action of the valve(s) and/or gate(s) can be facilitated by the controller168described above, or any other suitable element.

As noted above, in some embodiments, the system100can additionally or alternatively include a filter170, fluidly configured between the fluid channel outlet140and the manifold160, as shown inFIG. 1, that functions to prevent undesired substances from flowing into the fluid channel inlet120. The filter170preferably has a physical membrane that prevents substances having a governing dimension above a threshold size (e.g., defined by pores in the membrane); however, any other suitable mechanism can facilitate filtration of undesired substances from the fluid channel110. In one variation, the filter170can be configured immediately downstream of the fluid channel outlet140, in order to prevent undesired substances from entering the reservoir150. Additionally or alternatively, the system100can include a filter170configured within the reservoir, but upstream of the pump167, in order to prevent undesired substances from affecting proper function of the pump167and/or reaching the manifold160during recirculation of fluid into the fluid channel110. Additionally or alternatively, the filter170can be configured at any other suitable portion of a fluid loop defined across the manifold160, the fluid channel110, and the reservoir150. The filter170is preferably configured to be a replaceable element of the system110in order to promote ease of maintenance; however, the filter170can alternatively be configured in any other suitable manner. Variations of the system100can include a single filter, or can alternatively include multiple filters configured to provide redundancy in removing undesired substances from the fluid loop of the system100.

The system100can additionally or alternatively include a substrate actuation module190that transmits the substrate into the section-mounting region in a first operation, and delivers the substrate from the section-mounting region, with the section mounted to the substrate, in a second operation. The substrate actuation module190is configured to couple to an imaging substrate102, and functions to move the substrate relative to the section-mounting region130of the fluid channel110to facilitate placement of a section101onto the substrate102in an accurate and repeatable manner.

As shown inFIG. 6A, the substrate actuation module190can comprise a gripper191configured to couple to at least one surface106of a substrate102(e.g., glass slide), without obstructing mounting of a section101to the substrate, by any one or more of: friction, adhesion, compressive force, and any other suitable mechanism, in a manner that is consistent across all imaging substrates utilized by the system100. Furthermore, the substrate actuation module190preferably comprises an actuator192configured to induce motion of the gripper191and/or the imaging substrate along a path relative to a section at the section-mounting region130of the fluid channel. In one variation, the actuator192is a linear actuator configured to transmit the imaging substrate along a linear and sloping path into a reservoir of fluid defined at the section-mounting region130(e.g., immediately downstream of a section at the section-mounting region130), as described above; however, the actuator192can alternatively be configured to transmit the substrate102along any other suitable path that facilitates mounting of the section101onto the substrate102. The path along with the actuator192transmits the substrate102can be constrained by a rail193, as shown inFIG. 6Aor can alternatively be constrained or unconstrained in any other suitable manner.

Preferably, actuation in the substrate actuation module190is configured to coordinate with flow, from the fluid channel inlet120, to the section-mounting region130and out of the fluid channel outlet140. As such, the substrate actuation module190is preferably configured to cooperate with or be co-governed by the controller168of the pump167, in synchronizing flow of fluid through the system100and mounting of sections101to substrates102by way of the substrate actuation module190. In some variations, a flow rate into the fluid channel110can be reduced or halted by the controller168of the pump167to stabilize a position of the section101at the section-mounting region130prior to mounting; however, flow can be adjusted in any other suitable manner and with any suitable sequence that facilitates mounting of the section101to a substrate102.

In an example operation of the substrate actuation module190, as shown inFIGS. 13A-13C, the substrate actuation module190coordinates with flow into the fluid channel110as governed by the controller168of the pump167. In a first phase of the example operation, as shown inFIG. 13A, a section101bhas been transported to the section-mounting region130, as driven by fluid flow into the fluid channel110by the pump167. In the first phase of the example operation, flow is provided into the fluid channel110to bring the section101toward the section-mounting region130, with a substrate102partially submerged within the section-mounting region130by the substrate actuation module190. In the phase portion of the example operation, as shown inFIG. 13A, a section101ahas already been mounted to the substrate102, and an additional section101bis in position, at the section-mounting region130, to be mounted to the substrate102. In the state shown inFIG. 13Awith regions of the substrate102and section101defined inFIG. 14, the fluid level27in the fluid channel110is lower downstream of the substrate102than it is upstream of the substrate102, and the base surface15and sidewall14geometries of the fluid channel110at the section-mounting region130are configured to constrain the section101bto a desired lateral substrate position31. The sidewalls14of the fluid channel110, as shown inFIG. 6A, then widen around the substrate102at the section-mounting region130to provide a surface water velocity sufficient to maintain a position of the section102bintended to be mounted, even during flow speed reduction induced by the controller168. In the example operation, the sidewalls14are sufficiently close to the substrate105sides to provide enough construction, such that a drop in the fluid level27across the substrate102occurs during mounting of the section101bto the substrate102. In the first phase of the example operation, shown inFIGS. 13A and 14, the depth that the substrate102is submerged in the section-mounting region130establishes a line of juncture30between fluid in the section-mounting region130and the top of the substrate102, and therefore a vertical position29of the section101bbeing mounted to the substrate102.

In a second phase of the example operation, as shown inFIG. 13B, reducing the flow rate of fluid in the fluid channel110causes the section101bto be secured to the substrate102. An edge108of the section101bthat is in contact with the substrate102is the first portion of the section101bto be mounted to the slide, and as the fluid level equilibrates within the section-mounting region130, more of the section101bis mounted to the substrate102. In the second portion of the example operation, the entire section101bis mounted onto the substrate102prior to mechanical retraction of the substrate from the section-mounting region130by the substrate actuation module190; however, variations of the example operation can include any other suitable workflow that does not involve mounting of an entire section101bprior to retraction of the substrate102from the section-mounting region130. For instance, only a portion of the section101bcan be laid onto a substrate102by flow modulation in the fluid channel110, and mechanical retraction of the substrate102from the section-mounting region130by the substrate actuation module190consummates mounting of the section101bto the substrate by way of substrate withdrawal and an adhesion force produced by fluid between the section101band the substrate102. Alternatively, mechanical retraction of the substrate102from the section-mounting region130can cause application of the section101bto the substrate102substantially without modulation of a flow rate within the fluid channel110by the controller168of the pump167. Additionally or alternatively, modulation (e.g., lessening) of an angle of a substrate102within the section-mounting region130, by the substrate actuation module190, can be used to apply a section101bonto a substrate.

In a third portion of the example operation, as shown inFIG. 13C, retraction of the substrate102from the section-mounting region130provides a flow path (e.g., an unobstructed path) that allows undesired substances28(e.g., debris and discarded sections) to be removed by flowing out of the fluid channel outlet140, and optionally, through a filter170.

1.4 System—Temperature Regulation and Wrinkle Removal Module

As shown inFIG. 1, the system100can additionally or alternatively include a temperature regulating module180in contact with fluid from the reservoir, that adjusts a temperature of fluid within the fluid channel. As such, in facilitating mounting of sections at substrates, fluid from the reservoir150or portions of the fluid channel110can be transmitted at a desired temperature throughout the system. The desired temperature is preferably contained within a range of temperatures having a higher limiting temperature and a lower limiting temperature. The higher limiting temperature is preferably configured such that an embedding medium (e.g., paraffin wax) surrounding a specimen of a section101does not completely melt, and the lower limiting temperature is preferably configured such that the section101does not contract in a manner that could cause wrinkling or other damage of the section101.

In one variation, the temperature-regulating module180can be in communication with reservoir150in a manner that provides regulation of the temperature of fluid within the reservoir150, as it is transmitted from the reservoir150into the fluid channel inlet120. As such, temperature of the fluid at the reservoir150can be adjusted prior to delivery into the fluid channel110. Additionally or alternatively, the temperature-regulating module180can be in communication with any arbitrary position in the flow path of the fluid channel110to create a localized temperature profile at a desired portion of the fluid channel110, without requiring regulation of the entire volume of fluid in the reservoir150. In yet another alternative variation, the temperature-regulating element may induce indirect (e.g., non-contact) temperature variation of a section101at any point along flow through the fluid channel4(e.g., by air convection or radiant/infrared heating) without requiring direct thermal conduction between fluid in the fluid channel110or fluid at the reservoir1500, and a temperature-regulating module180. The reservoir150and/or the fluid channel110can, however, be configured in any other suitable manner.

The system100can additionally or alternatively include a wrinkle-removal module50, as shown inFIGS. 6 and 15, that functions to reduce or eliminate any wrinkling of sections prior to or during mounting to a substrate102. The wrinkle-removal module50can be configured proximal to the section-mounting region130of the fluid channel110, and functions to affect a local fluid parameter near a section in the section-mounting region130, such that the section101is substantially void of wrinkles prior to, during, and/or after coupling of the section101to a substrate102. The wrinkle-removal module50preferably modulates a local fluid temperature within the section-mounting region130, in coordination with delivery of the section101from the fluid channel inlet120to the section-mounting region130, as facilitated by the controller168of the pump167. As such, in a first variation, an example of which is shown inFIG. 16, the wrinkle-removal module50can include an injector51configured to inject a volume of fluid (e.g., from the reservoir, from another fluid source) into the fluid channel110proximal the section-mounting region130, wherein fluid from the injector51is at a temperature configured to increase fluidity of the section (e.g., a wax section) within the section-mounting region130. In this variation, the temperature of the fluid from the injector51is preferably elevated relative to a global fluid temperature within the fluid channel, to provide a local fluid temperature (e.g., to 40-60° C.) that increases fluidity of the section without complete dissociation or melting of the section. However, fluid can alternatively be provided from the injector51at any other suitable temperature that facilitates wrinkle removal in a section. In this variation, the system100can include a switch (e.g., 3-way switch) configured to switch between a first configuration in which fluid at a lower temperature from the reservoir150is circulated into the fluid channel110by way of the fluid channel inlet120, and a second configuration in which fluid at an elevated temperature (e.g., as passed through a heating apparatus upstream of the injector51) is circulated into the fluid channel110by way of the injector51. Additionally or alternatively, a flow rate used to deliver fluid at an elevated temperature from the injector51can be higher, lower, or substantially equal to a flow rate used to deliver fluid at a lower temperature into the fluid channel inlet120.

In a first example of the first variation, the injector51′ can be positioned superior to and upstream of the section-mounting region130, as shown inFIG. 17A, in order to inject high-temperature fluid into the fluid channel110upstream of the section-mounting region110, such that the high-temperature fluid flows under a section within the section mounting region130to remove any wrinkles in the section, prior to mounting of the section101to a substrate102. In a second example of the first variation, as shown inFIG. 17B, the injector51″ can be positioned downstream of the section-mounting region130and configured to inject high-temperature fluid upstream into the section-mounting region130, in order to remove any wrinkles in a section within the section-mounting region130. In a third example of the first variation, as shown inFIG. 17C, the injector51′″ can be positioned directly inferior to a section101within the section-mounting region130(e.g., at a base surface of the fluid channel110, within a reservoir into which the imaging substrate is directed for mounting of the section), such that high-temperature fluid is injected in an inferior-to-superior direction, toward the section101, to remove any wrinkles. In one variation of the third example, the fluid channel110can include a reservoir proximal the section-mounting region130, at which the section is held stationary and exposed to fluid at an elevated temperature, from the injector51, prior to mounting of the section to the imaging substrate. In another variation of the third example, a substrate102can be positioned (e.g., at an angle, perpendicularly) with an edge against the base surface of the fluid channel110proximal the section-mounting region130to create a dam, fluid at an elevated temperature can be delivered toward the imaging substrate from the injector51and trapped by the dam formed by the imaging substrate, and a section101positioned upstream of the dam can thus be positioned over a volume of fluid at an elevated temperature to undergo de-wrinkling. Then, the substrate can be positioned away from the base surface of the fluid channel4, thereby breaking the dam and allowing the section to be mounted to the imaging substrate, free of wrinkles, as the fluid level in the fluid channel110drops.

In a fourth example of the first variation, the injector51can be configured to deliver high-temperature fluid from sidewalls of the fluid channel110proximal section-mounting region130, in order to remove any wrinkles in a section within the section-mounting region. In a fifth example of the first variation, the injector51can be configured to deliver fluid at an elevated temperature through the same manifold160used to deliver fluid from the reservoir150into the fluid channel inlet150, in order to provide fluid at a suitable temperature for de-wrinkling of a section. In one variation of the fifth example, the entire volume of fluid from the reservoir150can be elevated to a desired temperature for de-wrinkling of a section, and delivered through the manifold160, by the injector, such that all fluid flowing within the fluid channel110is elevated to the desired temperature. In any of the above examples of the first variation, the section101can be held stationary by the substrate102or any other suitable object as fluid from the injector51flows under the section. Furthermore, a length of time over which the section101sits atop fluid at an elevated temperature can be adjusted according to requirements of a sample-type (e.g., tissue type) of the section, for instance, by adjusting flow rates of fluid into the fluid channel110and/or adjusting a position of the section by way of the substrate actuation module190. Variations of the injector51of the first variation can, however, be configured in any other suitable manner or implement combinations of any of the above examples/variations.

In a second variation of the wrinkle-removal module50involving local temperature adjustment, the wrinkle-removal module50can additionally or alternatively include a heating module52configured to provide convective and/or radiant heat transfer toward a section at the section-mounting region130. As shown inFIG. 18, the heating module52can be configured to transmit heat toward one or more surfaces of the section101, from a direction superior to and/or inferior to the section101. Furthermore, the heating module52can be configured to deliver heat toward the section prior to, during, and/or after contact between the section and a surface of a substrate102. As such, the heating module52can be configured to transmit heat toward the section101by locally heating fluid within the section-mounting region130and/or by transmitting heat through air toward a surface of the section101at the section-mounting region130, with or without a substrate102present. In a first example of the second variation, the heating module52can comprise a heating element positioned at a base surface of the section-mounting region130and inferior to a section at the section-mounting region130, such that the heating element locally heats fluid (e.g., by convective heat transfer) at an inferior surface of the section, thereby facilitating wrinkle removal. In a second example of the second variation, the heating module52can comprise heating elements spanning sidewalls of the fluid channel110proximal to (e.g., upstream of, adjacent to, downstream of, etc.) the section-mounting region130, such that the heating elements locally heat fluid (e.g., by convective heat transfer) from lateral directions to facilitate wrinkle removal. In a third example of the second variation, the heating module52can comprise a heating element positioned superior to a section at the section-mounting region130, configured to provide radiant and/or convective heat transfer through air toward the section at the section-mounting region130. In one variation of the third example, heated air from the heating module52can be delivered toward a substrate102with the section101, in order to heat the section101and residual fluid between the section and the imaging substrate to provide a de-wrinkling mechanism. Variations of the heating module52of the second variation can, however, be configured in any other suitable manner or implement combinations of any of the above examples/variations. Furthermore, delivery of heat from the heating module52to the section can be performed multiple directions simultaneously, and/or in any other suitable sequence of directions.

In any of the above examples and variations, the injector51/heating module52can be configured cyclically or non-cyclically vary temperatures proximal to a section within the section-mounting region130, in order to induce thermal expansion and contraction of the section101. In these variations, repeated expansion and/or contraction of the section can allow removal of any wrinkles that would remain after a single instance of heating of the section. Furthermore, in any of the above examples and variations, the injector51/heating module52can be configured to move (e.g., by coupling to an actuator) relative to a section101(e.g., by moving the injector/heating module, by moving an imaging substrate or the section relative to the injector/heating module) at the section-mounting region130, such that heat can be provided consistently to sections at the section-mounting region130in a dynamic manner. Moving and/or adjusting an angle of a substrate102with the section101relative to a heating module52can, for instance, facilitate wicking of fluid from the substrate102and facilitate drying of a section101during de-wrinkling, as shown inFIG. 19. Heating by the wrinkle-removal module50can furthermore be performed continuously as a stream of sections flow into the section-mounting region, or intermittently, for each section that flows in the section-mounting region.

In alternative variations, the wrinkle-removal module50can adjust local fluid behavior (e.g., flow behavior, viscosity behavior, etc.) proximal to a section within the section-mounting region130, in order to facilitate wrinkle removal within a section for mounting. In one alternative variation, the wrinkle-removal module50can generate uniformly or non-uniformly diverting flows proximal to (e.g., directly inferior to) a section at the section-mounting region130that provide forces that expand the section. In examples of this variation, the diverting flows can include two or more flow paths directed outward from a point proximal to a center point of a section101at the section-mounting region130. In another alternative variation, the wrinkle-removal module50can include an injector configured to transmit a fluid, different from fluid flowing from the fluid channel inlet120to the fluid channel outlet140, that provides an expanding force (e.g., based upon differences in density, based upon differences in viscosity, etc.) at a surface of a section101at the section-mounting region130. In another alternative variation, as shown inFIG. 20, the wrinkle-removal module50can include a vibration module53configured to generate vibration waves proximal to a section at the section-mounting region130, in order to facilitate wrinkle removal. In examples of this variation, the vibration module53can be configured to generate vibrations mechanically and/or acoustically, and can be configured to generate standing and/or non-standing waves. Variations of the wrinkle-removal module50can, however, comprise any other suitable elements configured to facilitate wrinkle removal by any other suitable mechanism.

In still alternative variations, the wrinkle removal module50can be configured to transfer heat to the substrate102, in order to increase the temperature of the substrate102to remove wrinkles in a section101that contacts the heated substrate102. As such, the wrinkle removal module50can comprise a heating element (e.g., heating plate, array of heating chips, etc.) configured to contact at least one surface of the substrate102and/or radiate heat toward the substrate102, as shown inFIG. 21, prior to or during mounting of the section101to the substrate102in order to remove wrinkles in the section101. In specific examples, the heating element can be integrated with the substrate actuation module190that is configured to manipulate motion of the substrate102, or can be configured to contact the substrate102in any other suitable manner.

Furthermore, the wrinkle-removal module50can be configured to coordinate with flow, from the fluid channel inlet120, to the section-mounting region130, as facilitated by the controller168of the pump167coupled to the manifold160. In some variations, a flow rate into the fluid channel110can be reduced or halted to stabilize a position of the section at the section-mounting region130, thereby facilitating wrinkle removal within the section101. Flow within the fluid channel110can, however, be configured in any other suitable manner in coordination with the wrinkle-removal module50.

In one example operation, involving coordination between flow governed by the controller168of the pump167, the wrinkle-removal module50, and the substrate actuation module190, the substrate actuation module190can be configured to transmit a substrate102to a desired fluid depth29, as shown inFIG. 22A, within the section-mounting region130of the fluid channel110. Then, a section101can be transmitted toward the section-mounting region130from the fluid channel inlet110after separation from a blade3of a microtome104of a sample-sectioning module103, as shown inFIG. 22B, and the wrinkle-removal module50can inject a volume of high-temperature fluid toward the section101to remove wrinkles, as shown inFIG. 22C. Flow from the fluid channel inlet120can then be increased to facilitate delivery of the section101onto the substrate102as the substrate102is retracted from the section-mounting region130by the substrate actuation module190, as shown inFIG. 22D. In variations involving mounting of multiple sections onto a single substrate102, the above example can be repeated multiple times, with the substrate102delivered into the section-mounting region130of the fluid channel110at successively decreasing depths for each section101mounted to the substrate102. Variations of the example can, however, involve any other suitable workflow.

1.5 System—Additional Elements and Alternative Configurations

Variations of the system100can alternatively omit any of the above described elements in order to provide simplified variations of the system100. For instance, one variation of the system100can include a reservoir150comprising fluid and configured to receive a section101at a surface of fluid in the reservoir150; and a wrinkle removal module50configured proximal to the section within the reservoir. In this simplified variation, the wrinkle removal module50can facilitate expansion of the section using one or more mechanisms as described above, and a human technician or other entity can mount the expanded section onto an imaging substrate manually or automatically. Variations of the above embodiments can further include any other suitable elements configured to facilitate mounting of a section onto an imaging substrate. For instance, a variation of the system100can include a tracking module configured to track a position of a section, and to facilitate wrinkle-removal within the section at any desired position in coordination with the tracking module. The system100can, however, omit and/or incorporate any other suitable element(s), or have alternative configurations, some of which are described below.

In one variation of the system100, one or more sensors75can be placed along the fluid channel110to enable detection of sections101as they pass, as shown inFIG. 23. The sensors can operate using any one or more of: infrared emitter-detector pairs, capacitive sensing, light contrast detection, image processing, and any other suitable mechanism to detect the presence, type, shape, and/or condition of a section101being transmitted through the fluid channel110. Such sensors can function to facilitate appropriate timing of flow modulation for placement of a section101onto a substrate102, as governed by a controller168of a pump167of the system100. Such sensors can additionally or alternatively enable detection of the presence or absence of fluid in the fluid channel110, a velocity of a section101as it is transmitted within the fluid channel110, physical parameters of (e.g., dimensions of, damage to, etc.) a section101within the fluid channel110, and any other suitable parameters. Such sensors can additionally or alternatively be used for subsequent, automated and/or manual, decision-making processes to adjust system performance, enable sorting of section, enable labeling or tagging of substrates with relevant information, and/or flagging conditions requiring operator intervention. The sensors can be mounted at an underside surface of the fluid channel110and configured to detect overhead passing sections (e.g., by way of transparent windows through the base surface of the fluid channel110), or can additionally or alternatively be mounted above the fluid channel110and configured to detect sections passing below.

To automate management of substrates for rapid exchange of substrates with mounted sections and empty substrates, a rail193of the substrate actuation module190can be mounted on a pivoting element197, as shown inFIG. 24. As such, the substrate actuation module190, with a gripper191mounted to a rail193can form a robotic arm that can be used for retrieving and replacing mounted substrates, for submerging substrates at the section-mounting region130during section placement, and for retraction to different positions for placement of multiple sections. To facilitate automated retrieval and replacement, the substrate actuation module193can interface with a substrate rack98(e.g., rack of imaging slides) having a set of slots (e.g., parallel slots, radially oriented slots, etc.) configured to hold substrates for retrieval and replacement. Substrate manipulation by the system100can, however, be automated in any other suitable manner.

To facilitate separation of adjoining sections produced by a sample sectioning module103, the system100can include elements that apply a treatment to the top or bottom of an embedded tissue section to create dissimilar materials at a section-section junction, thereby reducing the tendency of sections to wrinkle or form ribbons during processing. As shown inFIG. 25, the embedding medium90can comprise wax, and treatment92can comprise a quick-setting enamel applied to bottoms of sections101to prevent formation of ribbons or wrinkling.

In some variations, the microtome104of a sample sectioning module103interfacing with the system100can comprise a temperature-regulated chuck33, as shown inFIG. 24, that functions to maintain a bulk embedded sample at a desired temperature for sectioning. The temperature-regulated chuck33can also allow for consistent production of high-quality sections even if the bulk embedded sample is left within the chuck33for extended periods, such as when serial sectioning.

In some variations, the system100can include an atomizer198or other element, as shown inFIG. 24, configured to spray fluid over a sample to hydrate the bulk sample while it is being sectioned by the sample sectioning module103. The atomizer198can thus prevent the need for an operator to periodically remove, hydrate, and replace a sample during sectioning of specific types of tissues that are susceptible to dehydration and flaking during sectioning. Additionally or alternatively, the system100can include a permeable material (e.g., sponge, fabric, etc.), saturated with a hydrating fluid and configured to contact the bulk sample (e.g., by providing relative motion between the bulk sample and the permeable material), in order to prevent drying of the bulk sample. Hydration of the bulk sample to improve sample processing can, however, be performed in any other suitable manner.

In some variations, the system100can include a laser-etching device that functions to label substrates as they are mounted with sections, thus further reducing a need for operator intervention in substrate labeling. The laser-etching device can further function to reduce a possibility of mismatching substrate-mounted sections with their corresponding source embedded samples. The laser-etching device can be integrated with an informational technology (IT) system of a laboratory or clinic where the system100is in use.

In some variations, the system100can include an anti-static ionizing device that functions to ensure that each section101is electrostatically neutral at certain phases of processing. The anti-static ionizing device can minimize risk of electrostatic attraction or repelling of a cut section101toward a sidewall or other portion of the fluid channel110in a manner that could hinder mounting of the section to a substrate.

In some variations, the system100can include a kinetic sensor coupled to a blade3and/or sample mounting chuck of the sample sectioning module103that functions to sense acceleration, vibration, and/or any other kind of feedback that could be used to automatically adjust cutting motion of the sample sectioning module103.

The kinetic sensor can thus function to reduce operator interaction and improve automation in the system100.

In one alternative configuration, sections can be transmitted to a substrate from a path that is perpendicular to that described in the embodiments and variations above, which allows for a condensed fluid path. In another alternative configuration, as shown inFIG. 26, a linear flow boost (i.e., a burst of fluid flow) can be used to introduce flow around a submerged substrate102at the section-mounting region130to produce consistent section placement upon a substrate102.

The system100can, however, include any other suitable elements configured to facilitate mounting of one or more sections onto a substrate. Furthermore, as a person skilled in the art will recognize from the previous detailed description and from the figures and claims, modifications and changes can be made to the system100without departing from the scope of the system100.

As shown inFIG. 27, an embodiment of a method100for coupling a section to a substrate comprises: providing a fluid channel having a fluid channel inlet, a section-mounting region downstream of the fluid channel inlet, and a fluid channel outlet downstream of the section-mounting region S210; at the fluid channel inlet, receiving the section, processed by a sample sectioning module positioned proximal the fluid channel inlet S220; delivering the section from the fluid channel inlet toward the section-mounting region upon transmission of fluid flow into the fluid channel inlet, wherein transmission of fluid flow into the fluid channel inlet is governed by a controller S230; at a substrate actuation module, transmitting the substrate into the section-mounting region to receive the section, in a first operation, in coordination with the controller S240; and at the substrate actuation module, delivering the substrate, with the section mounted to the substrate, from the section-mounting region in a second operation, in coordination with the controller S250. In some embodiments, the method200can include any one or more of: transmitting fluid flow through the fluid channel outlet to be recycled into the fluid channel inlet, by way of a reservoir in fluid communication with the fluid channel inlet and outlet S260; and at a wrinkle-removal module proximal to the section-mounting region, transmitting heat toward the section, thereby mitigating wrinkling of the section at the substrate S270.

The method200functions to automate processing of sections (e.g., histological specimen sections, biological sections, etc.) in a manner that consistently generates high-quality mounted sections, with minimal or no effort from a human technician. As such, the method200can significantly reduce labor-intensive aspects of mounting sections to substrates. The method200is preferably implemented by at least a portion of the system100described in Section 1 above; however, the method200can additionally or alternatively be implemented using any other suitable system(s).

Block S210recites: providing a fluid channel having a fluid channel inlet, a section-mounting region downstream of the fluid channel inlet, and a fluid channel outlet downstream of the section-mounting region. Block S210functions to provide a fluid conveyer that can be used to drive a section for mounting at a substrate. Block S210is preferably implemented using an embodiment of the system100described above, and more specifically, using embodiments, variations, and/or examples of the fluid channel110, fluid channel inlet120, section-mounting region130, and fluid channel outlet140described above; however, Block S210can alternatively be implemented using any other suitable system100that provides a fluid path, with control of fluid flow parameters for automatically mounting histological sections to one or more substrates.

Block S220recites: at the fluid channel inlet, receiving the section, processed by a sample sectioning module positioned proximal the fluid channel inlet. Block S220functions to initiate sample reception within the fluid channel, such that the sample can be transmitted to downstream portions for manipulation (e.g., positioning, de-wrinkling) and mounting at a substrate. In Block S220, the section is preferably received from a bulk embedded sample processed by a sample sectioning module (e.g., comprising a microtome with a blade proximal the fluid channel inlet); however, the section can alternatively be received in Block S220in any other suitable manner. In some variations, Block S220can include one or more of: retaining an edge of the section by way of coupling the edge of the section to a cutting instrument of the sample sectioning module S222; and releasing the section into the fluid channel inlet upon separating the section from an adjoining section coupled to the cutting instrument of the sample sectioning module S224. Block S224, as described above, can include any one or more of: introducing fluid through a manifold into the fluid channel (e.g., at an angle Y) to free a preceding section for transmission into the fluid channel inlet S224a; generating fluid flow beneath a section within the fluid channel S224b, such that a shear force induced at a junction between sections provides separation; manually separating a section from the blade (e.g., using forceps) S224c; implementing an elevated floor of the fluid channel inlet, immediately downstream of the manifold, to cause fluid to be drawn away from the blade as it is delivered into the fluid channel; using a separation device (e.g., a paddle, a chuck, etc.), as shown inFIGS. 4A-4C, thereby providing a mechanical force that separates adjoined sections; and using any other suitable method of separating adjoining sections without damaging sections. Block S220can, however, include any other suitable steps for transmitting a section that has been cut from a bulk embedded sample into the fluid channel inlet.

Block S230recites: delivering the section from the fluid channel inlet toward the section-mounting region upon transmission of fluid flow into the fluid channel inlet, wherein transmission of fluid flow into the fluid channel inlet is governed by a controller. Block S230functions to drive a section, floating atop fluid within the fluid channel, toward the section-mounting region of the fluid channel upon transmission of fluid through a manifold in fluid communication with the fluid channel inlet. Block S230is preferably implemented using embodiments, variations, and/or examples of the fluid channel110, the pump167, the controller168, and the manifold160described in Section 1 above; however, Block S230can additionally or alternatively be implemented using any other suitable system or element(s). As shown inFIG. 29, delivering the section toward the section-mounting region in Block S230can thus include any one or more of: providing a progressively narrow fluid path within the fluid channel that enables accurate positioning the section onto a substrate within the section-mounting region S232; providing a descending fluid path within the fluid channel, thereby using gravity to facilitate acceleration of the section, atop a layer of fluid within the fluid channel, toward the section-mounting region S234; retaining a position of the section at the section-mounting region S236(e.g., upon modulation of fluid flow parameters); and using any other suitable block that enables accurate placement of a section at a substrate, in a repeatable manner.

Block S240recites: at a substrate actuation module, transmitting the substrate into the section-mounting region to receive the section, in a first operation, in coordination with the controller. Block S240functions to position a substrate at a desired depth and/or with a desired angle relative to a base surface of the fluid channel at the section-mounting region, which allows accurate positioning and mounting of the section to the substrate. Block S240is preferably implemented using an embodiment, variation, or example of the substrate actuation module190, substrate102, and section-mounting region130described in Section 1 above; however, Block S240can alternatively be implemented using any other suitable system or element(s). In variations, as shown inFIG. 30, Block S240can include any one or more of: transmitting a gripper of the substrate actuation module, with a substrate coupled to the gripper, along a path defined by a rail, wherein rail defines a sloping path into the section-mounting region and constrains motion of the substrate along the sloping path S242; defining a line of juncture between a portion of the substrate and fluid within the fluid channel at the section-mounting region S244; receiving the section at the line of juncture, thereby initiating mounting of the section to the substrate; modulating a fluid level at the section-mounting region, thereby promoting positioning and/or maintenance of a position of the section relative to the substrate S246; and performing any other suitable action that facilitates initial coupling of the section to the substrate. Block S240is thus preferably performed in coordination with modulation of flow parameters within the fluid channel by the controller, such that fluid parameters (e.g., flow velocity, flow acceleration, fluid level, etc.) for promoting accurate and repeatable coupling of a section to the substrate is substantially synchronized with motion of the substrate actuation module and coupled substrate.

Block S250recites: at the substrate actuation module, delivering the substrate, with the section mounted to the substrate, from the section-mounting region in a second operation, in coordination with the controller. Block S250functions to retract a substrate from a position within the section-mounting region, with a section at least partially coupled to the substrate, which allows the section to gradually be fully mounted to the substrate. Block S250is preferably implemented using an embodiment, variation, or example of the substrate actuation module190, substrate102, and section-mounting region130described in Section 1 above; however, Block S250can alternatively be implemented using any other suitable system or element(s). In variations, as shown inFIG. 31, Block S250can include any one or more of: retracting a gripper of the substrate actuation module, with a substrate coupled to the gripper, along a path defined by a rail, wherein rail defines a sloping path into the section-mounting region and constrains motion of the substrate along the sloping path S252; modulating a fluid level at the section-mounting region, thereby promoting mounting of the section onto the substrate S254by producing an adhesion force between the section and the substrate; and performing any other suitable action that facilitates initial coupling of the section to the substrate. Block S250is thus preferably performed in coordination with modulation of flow parameters within the fluid channel by the controller, such that fluid parameters (e.g., flow velocity, flow acceleration, fluid level, etc.) for promoting mounting of a section to the substrate in an accurate and repeatable manner is substantially synchronized with motion of the substrate actuation module and coupled substrate.

In some variations, Blocks S240and S250can be iteratively repeated for mounting of multiple sections to a single substrate, wherein a substrate is delivered to progressively decreasing depths within the section-mounting region to enable reception of multiple sections at desired positions along the substrate. An example workflow of mounting multiple sections to a single substrate is shown inFIGS. 13A-13C.

In some variations, the method200can include Block S260, which recites: transmitting fluid flow through the fluid channel outlet to be recycled into the fluid channel inlet, by way of a reservoir in fluid communication with the fluid channel inlet and the fluid channel outlet. Block S260functions minimize wasting of fluid by the system, by using recirculated fluid to process samples. Block S260is preferably implemented using embodiments, variations, and/or examples of the fluid channel outlet140, the reservoir150, the pump167, the controller168, the filter170, and the manifold160described in Section 1 above; however, Block S260can alternatively be implemented using any other suitable system or elements. Block S260preferably includes providing a flow path from the fluid channel to the reservoir, by way of the fluid channel outlet, and can additionally or alternatively include one or more of: filtering fluid at least at one of the fluid channel outlet, the reservoir, the pump, and the manifold S262, thereby removing undesired substances prior to recirculation of fluid through the system; modulating a temperature of fluid at least at one of the reservoir, the manifold, and a portion of the fluid channel S264; introducing an additive for surface tension modulation along with fluid from the reservoir, during recirculation S266; and any other suitable step that facilitates recycling of fluid in the system with minimal operator involvement.

In some variations, the method200can additionally or alternatively include Block S270, which recites: at a wrinkle-removal module proximal to the section-mounting region, transmitting heat toward the section, thereby mitigating wrinkling of the section at the substrate. Block S270functions to produce high-quality mounted sections, substantially free of wrinkling, upon transmitting heat to sections within the system100at desired stages of processing. Block S270is preferably implemented using an embodiment, variation, or example of the wrinkle-removal module50described in Section 1 above, whereby transmitting heat toward the section can include any one or more of: injecting fluid with a desired temperature toward a section at the section-mounting region S272(e.g., from underneath the section, from above the section, upstream of the section, downstream of the section, from sidewalls surrounding the section, etc); convectively transferring heat toward at least one surface of a section S274; heating a substrate to which a section is mounted or is intended to be mounted S276; and using any other suitable mechanism of heat transfer to de-wrinkle a section.

In one variation, as shown in FIGS.32and22A-22D, Block S270can include receiving the section at a fluid channel inlet of a fluid channel S310; delivering the section from the fluid channel inlet toward a section-mounting region of the fluid channel by way of fluid flow from a manifold proximal to the fluid channel inlet S320; delivering an imaging substrate into fluid at the section-mounting region at a first depth S330by way of a substrate actuation module; elevating a local temperature of fluid at the section-mounting region in coordination with delivery of the section into the section-mounting region S340to produce an expanded section; increasing flow into the section-mounting region, thereby delivering the expanded section toward the imaging substrate S350; and withdrawing the imaging substrate from the section-mounting region by way of the gripper module S360, thereby coupling the section to the imaging substrate.

However, Block S270can alternatively include removing wrinkles from a section prior to, during, or after mounting, using any other suitable apparatus. For instance, Block S270can include preventing wrinkling of a section by modulating a viscosity parameter or surface tension parameter of the fluid conveying the section to the section-mounting region, or by using acoustic vibrations to remove wrinkles from a section.

The method200can, however, include any other suitable blocks or steps configured to facilitate mounting of one or more sections onto a substrate in an automated or semi-automated manner. In one variation, the method200can include detecting, at a sensor system, one or more of: a section passing through a portion of the fluid channel, presence or absence of fluid in the fluid channel, a velocity of the section as it is transmitted within the fluid channel, physical parameters of (e.g., dimensions of, damage to, etc.) the section within the fluid channel110, and any other suitable parameters. In related variations, the method100can include timing flow modulation for placement of the section onto the substrate in response to signals generated by the sensor system.

In variations, the method200can additionally or alternatively include one or more of: applying a treatment to a portion of a bulk embedded sample used to generate the section, in order to prevent section wrinkling; automatically regulating a temperature of the bulk embedded sample; automatically spraying fluid over a portion of the bulk embedded sample to hydrate the sample while it is being sectioned by a sample sectioning module; with a laser-etching device, automatically labeling substrates as they are mounted with sections; with an anti-static ionizing device, ensuring that the section is at an electrostatically neutral state during processing; based upon signals from a kinetic sensor coupled to the sample-sectioning module, automatically modulating sectioning parameters to improve section quality; and any other suitable step that automates sample processing and/or improves quality of samples.

Variations of the system100and method200include any combination or permutation of the described components and processes. Furthermore, various processes of the preferred method can be embodied and/or implemented at least in part as a machine configured to receive a computer-readable medium storing computer-readable instructions. The instructions are preferably executed by computer-executable components preferably integrated with a system and one or more portions of the control module155and/or a processor. The computer-readable medium can be stored on any suitable computer readable media such as RAMs, ROMs, flash memory, EEPROMs, optical devices (CD or DVD), hard drives, floppy drives, or any suitable device. The computer-executable component is preferably a general or application specific processor, but any suitable dedicated hardware device or hardware/firmware combination device can additionally or alternatively execute the instructions.