U.S. patent application number 14/631586 was filed with the patent office on 2015-06-18 for device to collect and segregate tissue samples sectioned on a microtome.
The applicant listed for this patent is Vertex Pharmaceuticals, Inc.. Invention is credited to Jorge Arturo Ayala, Jack Barger, Michael Paul Emery, Sanjay Shivayogi Magavi, James Edward Sinclair.
Application Number | 20150168277 14/631586 |
Document ID | / |
Family ID | 46604057 |
Filed Date | 2015-06-18 |
United States Patent
Application |
20150168277 |
Kind Code |
A1 |
Magavi; Sanjay Shivayogi ;
et al. |
June 18, 2015 |
DEVICE TO COLLECT AND SEGREGATE TISSUE SAMPLES SECTIONED ON A
MICROTOME
Abstract
Disclosed herein are apparatuses and methods handling a portion
of a tissue sample when sectioned by a microtome. The apparatuses
include a container, a tissue sample holder in the container, and
one or more outlets configured to allow flow of a fluid from the
container. the flow through the outlet causes a portion of the
tissue sample sectioned by a microtome to move into the outlet. The
methods include sectioning one or more portions of a tissue sample,
and flowing a fluid past the tissue sample to cause the one or more
portions of the tissue sample to move away from the tissue sample
and toward at least one fluid outlet.
Inventors: |
Magavi; Sanjay Shivayogi;
(Cambridge, MA) ; Barger; Jack; (Poway, CA)
; Emery; Michael Paul; (Santee, CA) ; Sinclair;
James Edward; (Carlsbad, CA) ; Ayala; Jorge
Arturo; (San Diego, CA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Vertex Pharmaceuticals, Inc. |
Cambridge |
MA |
US |
|
|
Family ID: |
46604057 |
Appl. No.: |
14/631586 |
Filed: |
February 25, 2015 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
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13187416 |
Jul 20, 2011 |
8967024 |
|
|
14631586 |
|
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Current U.S.
Class: |
435/30 ;
435/309.1 |
Current CPC
Class: |
Y10T 83/222 20150401;
Y10T 83/0453 20150401; Y10T 83/2216 20150401; G01N 1/312 20130101;
Y10T 83/2066 20150401; Y10T 83/2179 20150401; G01N 1/06
20130101 |
International
Class: |
G01N 1/31 20060101
G01N001/31 |
Claims
1. An apparatus for handling a portion of a tissue sample when
sectioned by a microtome, the apparatus comprising: a container; a
tissue sample holder in the container; and an outlet configured to
allow flow of a fluid from the container; wherein the outlet is
positioned proximate to the tissue sample holder such that flow
through the outlet causes a portion of a tissue sample sectioned by
a microtome to move into the outlet.
2. The apparatus of claim 1, further comprising a microtome, the
microtome including a control system configured to automate a
portion of the microtome.
3. The apparatus of claim 1, wherein the tissue sample holder is
removable from the container.
4. The apparatus of claim 1, wherein the tissue sample holder
comprises at least two separable portions.
5. The apparatus of claim 1, comprising a first outlet and a second
outlet positioned proximate to the tissue sample holder such that
flow through the first outlet causes a portion of a first tissue
sample sectioned by a microtome to flow into the first outlet, and
flow through the second outlet causes a portion of a second tissue
sample sectioned by a microtome to flow into the second outlet.
6. The apparatus of claim 1, comprising a drain positioned in the
container.
7. The apparatus of claim 1, comprising a valve in fluid
communication with at least one of the one or more outlets and
configured to control the flow of fluid from the outlet.
8. The apparatus of claim 1, comprising a first conduit in fluid
communication with at least one of the one or more outlets.
9. The apparatus of claim 8, comprising a second conduit in fluid
communication with the outlet.
10. The apparatus of claim 9, wherein the second conduit is in
fluid communication with the outlet at a connection point located
in the first conduit, and further comprising a valve configured to
control the flow of fluid from the first conduit, the valve located
downstream of the connection point.
11. The apparatus of claim 10, further comprising a filter
positioned to prevent the portion of the tissue sample from flowing
into the second conduit.
12. The apparatus of claim 1, further comprising a storage plate,
the storage plate including one or more wells configured to receive
the one or more portions of the tissue sample.
13. The apparatus of claim 12, further comprising a moveable tray
configured to hold one or more storage plates with respect to the
one or more outlets.
14. The apparatus of claim 12, wherein one or more of the wells
comprises one or more apertures to allow fluid to flow through a
portion of the well without allowing a tissue sample section to
flow through the apertures.
15. The apparatus of claim 1, wherein the cross-section of at least
one of the one or more outlets is non-round.
16. A method of handling one or more portions of a tissue sample
sectioned by a microtome, the method comprising the steps of:
sectioning one or more portions of a tissue sample; and flowing a
fluid past the tissue sample to cause the one or more portions of
the tissue sample to move away from the tissue sample and toward at
least one fluid outlet.
17. The method of claim 16, wherein sectioning the one or more
portions of a tissue sample comprises sectioning a plurality of
portions of a tissue sample with a microtome.
18. The method of claim 16, wherein sectioning one or more portions
of a tissue sample comprises sectioning a portion of a plurality of
tissue samples with a microtome at substantially the same time.
19. The method of claim 16, wherein sectioning one or more portions
of a tissue sample comprises sectioning a first portion of a tissue
sample and a second portion of a tissue sample, wherein the second
portion of the tissue sample is sectioned sequentially with respect
to the first portion.
20. The method of claim 16, wherein flowing the fluid comprises
increasing flow through the at least one outlet in response to
completing a cutting stroke during the sectioning.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] Any and all applications for which a foreign or domestic
priority claim is identified in the Application Data Sheet as filed
with the present application are hereby incorporated by reference
under 37 CFR 1.57.
[0002] This application is a continuation of U.S. patent
application Ser. No. 13/187,416, filed Jul. 20, 2011, (now issued
as U.S. Pat. No. 8,967,024 to Magavi et al.), the disclosures of
which are incorporated herein by reference in their entirety.
BACKGROUND OF THE INVENTION
[0003] 1. Technical Field
[0004] The embodiments disclosed herein relate to microtome systems
for sectioning tissue samples.
[0005] 2. Introduction
[0006] A microtome apparatus is one example of a sectioning
instrument used extensively to cut extremely thin sections of
material, including, for example, plant tissue, animal tissue, or
other types of tissue samples. Conventional microtomes, such as
microtome 50 shown in FIG. 1, include a blade 51 that can be
extended to cut off a single, thin section or portion 71 from the
top of a tissue sample 70 supported on a tissue support structure
30 while submerged in a reservoir in a container 20. After
sectioning, the portion 71 floats in the reservoir (typically
aqueous buffer solution, or the like) within a container 20, and is
subsequently manually transferred with a tool such as a swab or
tweezers to a container such as a well of a multi-well plate, a
Petri dish, a microscope slide, or the like, to allow for further
analysis, testing and/or storage of the section.
[0007] Conventional microtome processes can be repetitive, tedious
and laborious, requiring dedicated laboratory personnel to
continuously operate and monitor the apparatus as each section is
produced. Further, the tissue samples can become contaminated or
damaged during their handling by laboratory personnel, increasing
operational costs. Thus, there is a need for an efficient microtome
apparatus and process that can provide high quality, sectioned
tissue samples with reduced waste and contamination.
SUMMARY OF THE INVENTION
[0008] In one embodiment, an apparatus for handling a portion of a
tissue sample when sectioned by a microtome is provided. The
apparatus includes a container, a tissue sample holder in the
container, and one or more outlets configured to allow flow of a
fluid from the container. The one or more outlets is positioned
proximate to the tissue sample holder such that flow through the
outlet causes a portion of the tissue sample sectioned by a
microtome to move into the outlet.
[0009] In another embodiment, a method of handling one or more
portions of a tissue sample sectioned by a microtome is provided.
The method comprises the steps of sectioning one or more portions
of a tissue sample, and flowing a fluid past the tissue sample to
cause the one or more portions of the tissue sample to move away
from the tissue sample and toward at least one fluid outlet.
[0010] In yet another embodiment, an apparatus for handling one or
more portions of a tissue sample sectioned by a microtome is
provided. The apparatus includes means for sectioning at least one
portion of a tissue sample and means for flowing a fluid past the
tissue sample. The means for flowing the fluid past the tissue
sample cause the one or more portions of the tissue sample to move
away from the tissue sample and toward at least one fluid
outlet.
[0011] For purposes of summarizing the invention and the advantages
achieved over the prior art, certain objects and advantages of the
invention have been described above and as further described below.
Of course, it is to be understood that not necessarily all such
objects or advantages may be achieved in accordance with any
particular embodiment of the invention. Thus, for example, those
skilled in the art will recognize that the invention may be
embodied or carried out in a manner that achieves or optimizes one
advantage or group of advantages as taught herein without
necessarily achieving other objects or advantages as may be taught
or suggested herein.
[0012] All of these embodiments are intended to be within the scope
of the invention herein disclosed. These and other embodiments of
the present invention will become readily apparent to those skilled
in the art from the following detailed description of the preferred
embodiments having reference to the attached figures, the invention
not being limited to any particular preferred embodiment(s)
disclosed.
BRIEF DESCRIPTION OF THE DRAWINGS
[0013] These and other aspects of the invention will be readily
apparent from the following description and from the appended
drawings (not necessarily to scale), which are meant to illustrate
and not to limit the invention, and in which:
[0014] FIG. 1 is a side view of a conventional apparatus for
sectioning a portion of a tissue sample.
[0015] FIG. 2 is a side view of one embodiment of an apparatus for
handling a portion of a tissue sample.
[0016] FIG. 3 is a flowchart illustrating a method of handling one
or more portions of a tissue sample sectioned by a microtome,
according to one embodiment.
[0017] FIG. 4 is a perspective view of one embodiment of an
apparatus for handling a portion of a tissue sample, with a portion
of the view shown as a cross-section taken along line 4-4 of FIG.
5.
[0018] FIG. 5 is a partial exploded perspective view of one
embodiment of the apparatus of FIG. 4.
[0019] FIG. 6 is a side cross-sectional view of one embodiment of
an apparatus for handling a portion of a tissue sample taken along
line 6-6 of FIG. 4.
[0020] FIG. 7 is a perspective view of one embodiment of a storage
plate configured to receive one or more portions of a tissue
sample.
[0021] FIG. 8 is a perspective view of one embodiment of the
apparatus for handling a portion of a tissue sample of FIG. 4, with
some components not shown for clarity.
DETAILED DESCRIPTION
[0022] The embodiments disclosed herein relate to apparatus and
methods for handling a portion of a tissue sample when sectioned by
a microtome.
[0023] Embodiments will now be described with reference to the
accompanying Figures, wherein like numerals refer to like elements
throughout. The terminology used in the description presented
herein is not intended to be interpreted in any limited or
restrictive manner, simply because it is being utilized in
conjunction with a detailed description of certain specific
embodiments disclosed herein. Furthermore, embodiments disclosed
herein may include several novel features, no single one of which
is solely responsible for its desirable attributes or which is
essential to the embodiments herein described.
[0024] FIG. 2 is a side view of one embodiment of an apparatus 10
for handling a portion 71 of a tissue sample 70. FIG. 3 is a
flowchart illustrating a method of handling one or more portions of
a tissue sample sectioned by a microtome, according to one
embodiment. It will be understood that the actions summarized in
the flowchart in FIG. 3 are neither exhaustive nor exclusive, and
that additional actions may intervene between those disclosed.
Furthermore, not all of the disclosed actions must occur. While the
following description refers to the apparatus of FIG. 2, it will be
appreciated that the method of FIG. 3 can be applied to other
apparatuses disclosed herein, as well as to other suitable tissue
sample handling apparatuses.
[0025] Referring to FIGS. 2-3, one or more portions 71 of tissue
sample 70 can be sectioned (block 12 of FIG. 3) from tissue sample
70. Fluid can then be flowed (block 14 of FIG. 3) past the tissue
sample 70 to cause the one or more portions 71 of the tissue sample
70 to move away from the tissue sample 70 and toward at least one
fluid outlet 40. For example, as described elsewhere herein, fluid
can be flowed (in some embodiments, using a laminar or nearly
laminar flow) from an inlet to a drain (e.g., FIG. 4) within
container 20, to move flow generally towards and past the tissue
sample 70. After entering the fluid outlet, the sectioned slice 71
of the tissue sample can be routed along a fluid channel to a
designated container such as a specific well of a multiwell plate.
As will be explained further below, as sections of the sample are
made, they can sequentially move out the outlet and into the
channel under the influence of the flowing fluid, and into
successive desired containers. The movement of a set of containers
or movement of the end of the channel can also be automated, so
that sectioning itself and the transfer/registration of the
sectioned samples can occur without manual manipulation of any
samples or containers, freeing laboratory personnel of this
previous time consuming manual task.
[0026] In some embodiments, flowing the fluid 14 can comprise
increasing flow through the at least one outlet 40 in response to
completing a cutting stroke during the sectioning 12. Such flow
increase can be provided, for example, by moving a valve to an open
position at the point when the cutting stroke is completed. The
valve can be opened for various lengths of time; in one embodiment,
the valve is opened for approximately 400 ms. In some embodiments,
one or more sensors (e.g., FIG. 6) can provide feedback as to when
the cutting stroke is completed, or near completion, to control the
timing of the opening and closing of valve.
[0027] In some embodiments, flowing the fluid 14 comprises allowing
flow through the one or more outlets 40 at a first flow rate prior
to the completing a cutting stroke, and allowing flow through the
one or more outlets at a second larger flow rate subsequent to
completing a cutting stroke. The first flow rate can be zero, or
can be a relatively small bleed flow. The second flow rate can
correspond to increasing flow through the one or more outlets 40,
for example, by moving a valve to an open position.
[0028] In some embodiments, the method of handling one or more
portions of a tissue sample sectioned by a microtome can further
comprise inhibiting the flow of a fluid through the one or more
outlets 40. Inhibiting can comprise restricting the flow by closing
a valve in the outlet channel.
[0029] Sectioning 12 can comprise sectioning a plurality of
portions 71 of a tissue sample 70 with a microtome 50. Such
sectioning a plurality of portions 71 of a tissue sample 70 can
comprise sectioning a first portion and a second portion, either
sequentially, or at substantially the same time.
[0030] For example, blade 51 of microtome 50 can be positioned with
respect to the tissue sample 70 such that the top of tissue sample
70 is slightly above (e.g. 10, 50, 100 micrometers, for example)
the cutting plane of blade 51. Blade 51 can advance, (e.g., in the
direction shown by directional arrow 502; FIG. 4), to section a
first portion 71 from tissue sample 70. Blade 51 can be advanced at
a variety of speeds. The blade speed and flow rates during
sectioning are preferably selected to facilitate clean cutting of
the section and smooth movement of the sample toward the outlet 40
and into the channel. Flow rate and blade speed during the cutting
process may be selected to cause the portion 71 of tissue sample 70
that is being sectioned to curl upward away from the tissue sample
70 and toward the outlet 40 as the cutting progresses. However,
blade speeds and flow rates that are too large can cause the
section 71 to fold over onto the front of the tissue sample 70.
This is preferably avoided, as it may cause the blade 51 to ride up
and over the folded section 71, which can result in an incomplete
cut and retention of the section 71 to the front of the tissue
sample 70. As mentioned above, shortly after the cut is complete,
the flow rate through the outlet 40 may be increased to wash the
section 71 into the outlet 40 and out the channel. After the first
portion is sectioned from sample 70, blade 51 can retract (e.g., in
the direction shown by directional arrow 502; FIG. 4).
[0031] Tissue sample 70 and blade 50 can then be positioned with
respect to each other, by moving sample 70 and/or blade 50 (e.g.,
in either or both of directions 503 and 504, respectively; FIG. 4),
such that a second portion of tissue sample 70 is above the cutting
plane of blade 51. The flow rate may then be decreased (or possibly
stopped completely), and blade 51 can again be advanced, to section
a second portion of tissue sample 70 with blade 51. In this way, a
plurality of portions of a tissue sample 70 can be sectioned with
microtome 50, wherein the second portion of the sample is sectioned
sequentially with respect to the first portion. The aforementioned
sequential sectioning operation can be repeated, for example, until
the tissue sample 70 is depleted, and/or replaced with another
sample. As sections are completed, each one can be routed to a
desired container in an automated manner.
[0032] In some embodiments, two or more tissue samples 70 can be
mounted on tissue sample holder 30. As shown in more detail below,
each tissue sample may be associated with its own outlet. Blade 51
of microtome 50 can be positioned with respect to the two or more
tissue samples 70 such that a first portion of a first tissue
sample 70 and a second portion of a second tissue sample 70 are
above the cutting plane of blade 51. Blade 51 can be advanced to
section the first and second portions 71 from the first and second
tissue samples 70, for example, in a single cutting stroke. In this
way, sectioning a plurality of portions 71 of a tissue sample 70
with microtome 50 can comprise sectioning a plurality of portions
71 of a plurality of tissue samples 70 with microtome 50 at
substantially the same time. This operation for sectioning a
plurality of portions of a plurality of tissue samples 70 can be
repeated, for example, using the sequential operation described
above, to sequentially section two or more portions of each tissue
sample in the plurality of tissue samples.
[0033] As used herein, "substantially the same time" means that a
single cutting stroke of blade 51 need not necessarily section the
two or more tissue samples 70 simultaneously. For example, the edge
of blade 51 can comprise an irregular shape, such that a first
portion of blade 51 cuts a first tissue sample prior to, but at
substantially the same time as, a second portion of blade 51
cutting a second tissue sample, during the same cutting stroke.
Additionally, the orientation of the edge of blade 51 and the two
or more samples 70 can be substantially parallel or non-parallel
with respect to each other, such that a single cutting stroke cuts
through two or more tissue samples sequentially.
[0034] FIGS. 4-6 illustrate various views of one embodiment of one
specific apparatus 10 for handling a portion 71 (e.g., a sectioned
layer or slice) of one or more tissue samples 70 when a sample 70
is sectioned by a microtome 50 that has been fabricated by the
applicants. Apparatus 10 can include a container 20, and a tissue
sample holder 30 configured to support the one or more tissue
samples 70. FIG. 4 shows a cutaway view of the front of the tray of
FIG. 5 to illustrate the outlets 40 and the coupling between the
outlets and the channels that route the tissue sections to their
desired containers.
[0035] Apparatus 10 can include one or more outlets 40 configured
to allow flow of a fluid from the container 20. The outlets 40 can
be positioned proximate to the tissue sample holder 30 such that
flow through the outlet 40 causes a portion 71 of the tissue sample
70 sectioned by the microtome 50 to move into the outlet 40. One or
more conduits (e.g., conduits 41, 42; FIGS. 1 and 3) may be
configured to be in fluid communication with outlet 40, as
described further herein. In some embodiments, apparatus 10 can
include one or more storage plates 90 (FIGS. 1 and 5) configured to
receive and store the sectioned portion 71 of the tissue sample 70.
In some embodiments, apparatus 10 can include a tray (e.g., a
movable tray) 110 (FIGS. 1 and 4) that can support the storage
plate 90.
[0036] Microtome 50 can comprise any of a number of different
microtome devices known or described herein. Microtome 50, and many
of the components of apparatus 10 described herein, can be manually
and/or automatically operated, such as with a control system and/or
other automation components. Apparatus 10 can include one or more
movable elements that comprise any of a number of devices that can
facilitate linear and/or rotational motion between two components.
Additionally or alternatively, many of the fluid control components
described herein, such as the valves, drains, flow controllers,
etc., can be automatically or manually controlled.
[0037] Microtome 50 can comprise a sectioning element (e.g., a
knife or blade) 51 attached to an arm or other blade support
structure 53 that can advance (e.g., directional arrow 502) and
retract (e.g., directional arrow 501) with respect to one or more
tissue samples 70, such that blade 51 can section a portion 71 of
tissue sample 70. In some embodiments, the blade 51 and the one or
more tissue samples 70 can move with respect to each other (e.g.,
directional arrows 503; 504, or approximately vertically), to allow
two or more portions 71 to be sequentially sectioned from a tissue
sample 70, or to adjust the thickness of the portion 71 of the
tissue sample 70 that is sectioned. For example, blade 51 can be
held stationary, and tissue sample 70 can be moved slightly
upwardly (e.g., approximately 50 .mu.m), to section a portion 71
from sample 70.
[0038] In some embodiments, blade 51 can move laterally with
respect to tissue sample 70 (e.g., directional arrows 505-506), for
example, to select the portion of the blade 51 that sections one or
more tissue samples 70 during the cutting stroke. Any of the
relative movement (e.g., in any of directions 501-506) between
blade 51 and one or more tissue samples 70 can be provided by
attaching one or more of the aforementioned movable elements to
blade 51 and tissue samples 70, or an intermediary structure, such
as tissue sample holder 30, container 20, arm 53, or other
intermediary structure. In a preferred embodiment, microtome 50 is
a vibrating microtome, and thus is configured to vibrate blade 51
during the sectioning of tissue samples 70, to provide a cleaner
and more accurate cut.
[0039] Referring to FIG. 4, apparatus 10 can include a controller
or processor element that can be implemented to automate and/or
control various aspects of the apparatus 10 and methods of using
apparatus 10 described herein. It will be appreciated that the
processor element of the apparatus 10 may be integral to the
housing of the apparatus 10, or all or part of the processing and
control circuits can be separate from the apparatus 10 itself. In
some embodiments, the processor can be a specialized
microcontroller which is designed specifically for controlling the
elements of the apparatus 10. Alternatively, the processor can be a
standard personal computer device such as an Intel processor-based
PC running an off the shelf operating system such as Windows,
Linux, MacOS, or the like. As used herein, the term "processor"
generally refers to one or more logic and control circuits which
are connected to the apparatus 10 to control the operation of
various components of the apparatus as described herein. In some
embodiments, the processor can include direct hardware interface
such as a USB port, an RS232 interface, and IP network interface
(wired or wireless), or some other type of connection, to load
software to control the components and functions of the apparatus
10. In some embodiments, the processor is integrated into the
apparatus, which then interfaces with a touch-screen user interface
that enables the user to set the parameters for automated control
of the different components of the apparatus.
[0040] In some embodiments, the processor can include software that
allows the user to enter the timing and parameters for controlling
one or more components of the apparatus 10, such as the movement of
microtome 50, container 20, storage plate 90, and/or tray 110 with
respect to each other, and the control of the flow of fluid within
and/or through any of these components. In some embodiments, the
software allows the user to program the apparatus to complete a
procedure for sectioning one or more tissue sample(s), including a
first sectioning step (e.g., the movement of the microtome blade to
section a tissue sample), the flow of fluid and movement of the
sectioned tissue sample into the outlet, the movement of the
sectioned tissue sample into a storage tray, and the indexing of
the storage tray prior to repeating these steps on a second tissue
sample section. In some embodiments, the processor can allow for
automated collection of "run data" including, for example,
temperature, pressure, flow and volume measurements, count of
sectioned tissue samples, operator identity, date and time,
etc.
[0041] Continuing to refer to FIGS. 4-6, tissue sample holder 30
can comprise any of a number of shapes and materials capable of
supporting one or more tissue samples 70 within container 20 in a
position to facilitate the sectioning of samples 70 by a microtome.
Tissue sample holder 30 can be a rigid or semi-rigid material, such
as metal, glass, ceramic, or plastic, and can be a flexible or
semi-flexible material, such as plastic. Tissue sample holder 30
and its components generally comprise a material with sufficient
rigidity and strength to support tissue sample 70 when being
sectioned by microtome 50. Tissue sample holder 30 can also
comprise a shape and material that is reasonably smooth and free
from burrs or sharp edges, to facilitate low turbulence fluid flow
on and around its surfaces, and to prevent snagging or catching of
objects on its surface. Tissue sample holder 30 preferably
comprises a hydrophobic material, and more preferably, a material
suitable for use within a clinical environment for handling tissue
samples. Tissue sample holder 30 can comprise any combination of,
and/or can be coated with, one or more of the aforementioned
materials.
[0042] Tissue sample holder 30 can be attached (e.g., permanently
or removably) to a portion of container 20, or an intervening
structure. Tissue sample holder 30 can comprise a separate
component, or can be formed integrally with container 20. For
example, tissue sample holder 30 can comprise a portion of a
surface of base 22 of container 20, or a surface of a structure
protruding from base 22, on which tissue sample 70 can be
positioned and sectioned by microtome 50. It will be understood
that tissue sample holder 30 can be configured to support any of a
number of different quantities of tissue samples 70, and is shown
supporting four tissue samples 70 for illustrative purposes only.
Moreover, the tissue samples 70 are shown in a single row, to be
sectioned by a single microtome blade 51 for illustrative purposes
only. Embodiments of apparatus 10 can include additional rows
and/or columns of tissue samples, to form various 2 or
3-dimensional arrays or matrices of tissue samples, to be sectioned
by two or more microtome blades at various orientations, either
simultaneously or sequentially.
[0043] Tissue sample holder 30 can comprise a tissue sample holder
base 31 configured to support one or more tissue samples 70. Base
31 can be any of a number of shapes, such as an elliptical,
rectangular, trapezoidal, or other regular or irregular shape. Base
31 can include an upper surface 31a which can face towards and
support tissue sample 70, and a lower surface 31b, which can face
towards and be supported by container 20 (e.g., base 22 of
container 20) when tissue sample holder 30 is attached to container
20 or an intermediate structure.
[0044] Tissue sample holder 30 can include one or more handling
portions configured to assist in the handling of tissue sample
holder 30. For example, the handling portion(s) can facilitate the
deployment and removal of tissue sample holder 30 to and from
container 20 with embodiments in which tissue sample holder 30 is
removably attached to container 20. The handling portions can
include one or more knobs, dimples, nipples, surface textures and
contours (e.g., convex protrusions and concave recessions), ribs,
slots, grooves, arms, tabs, hooks, handles, and the like, spanning
across and/or extending from various portions of sample tissue
sample holder 30. These handling portions can comprise one or more
materials that may improve a user's grip, such as various textured
frictional coatings, or resilient materials, such as rubber or
foam.
[0045] Referring to FIG. 5, tissue sample holder 30 can include one
or more handling portion(s) comprising one or more arms 32
extending from a portion of base 31 (e.g., from its opposed ends)
to facilitate the handling of tissue sample holder 30. Arms 32 can
include a lower portion 32a attached to base 31, and an upper
portion 32b that is attached to and extends from a portion of lower
portion 32a (e.g., a distal end). Portions 32a, 32b can extend in a
number of directions and/or at various angles with respect to each
other and/or base 31 (and/or with respect to a portion of container
22 (e.g., base 22) when tissue sample holder 30 is attached
thereto). Lower portion 32a can extend upwardly (e.g. orthogonally)
with respect to upper surface 31a of base 31. Upper portion 32b
preferably extends outwardly from lower portion 32a, to further
facilitate handling of tissue sample holder 30. In some
embodiments, lower portion 32a and upper portion 32b are of a
sufficient length and configuration with respect to base 31 and
container 20, such that tissue sample holder 30 can be deployed
and/or removed from container 20 without draining the fluid from
container 20, and without submersing upper portion 32b into the
fluid. Such an embodiment can allow a user to attach and detach
tissue sample 30 to and from container 20 without contaminating or
contacting the fluid within container 20.
[0046] Tissue sample holder 30 can be attached to container 20 with
sufficient strength and dimensional accuracy to support and align
tissue sample holder 30 with respect to microtome 50, thus
providing accurate and reliable sectioning of the portion(s) of
sample(s) 70 with microtome 50 described herein. A strong and
accurate alignment between tissue sample holder 30 and container 20
can also improve the alignment between tissue sample holder 30 and
outlet(s) 40. Such alignment can improve the accuracy and
reliability of the movement of the portion(s) of sample(s) 70 into
outlet(s) 40 when they are sectioned by microtome 50.
[0047] Tissue sample holder 30 and container 20 can be attached,
and preferably, removably attached, to each other with one or more
engagement elements. The engagement elements described herein can
comprise any of a variety of engagement and attachment structures
and methods, including adhesive, snaps, hooks, tabs, buttons, a
press fit, interference fit, snap fit, slots, grooves, screws,
rivets, and the like. The engagement elements can be attached to
and positioned along any of a number of portions of tissue sample
holder 30 and container 20, such as base 31 (e.g., bottom surface
31b), arms 32, and/or base 22. In the illustrated embodiment,
tissue sample holder 30 can include one or more tabs 33, extending
from a portion of one or more arms 32. Tabs 33 can be configured to
removably engage with a corresponding structure, such as a recess,
slot or groove, within container 20 (e.g., on base 22 of container
20).
[0048] In some embodiments, tissue sample 70 can be attached
directly to a portion of tissue sample holder 30, such as base 31
(e.g., upper surface 31a). In other embodiments, tissue sample 70
can be attached to an optional intermediary structure positioned
between tissue sample 70 and tissue sample holder 30. Tissue sample
70 can be attached to tissue sample holder 30 or an intermediary
structure using sutures, stitching, tissue adhesive (e.g.,
cyanoacrylate, etc.) or any suitable methods or structures known or
described herein for attaching tissue samples to a plastic or other
material used for tissue sample holder 30 or an intermediary
structure.
[0049] It may be a challenge to mount and/or remove tissue
sample(s) 70 onto tissue sample holder 30, as the available space
between a tissue sample 70 mounted on base 31 and another, adjacent
portion of tissue sample holder 30 (e.g., arms 32) may be limited,
making the mounting and removal procedure difficult, even with
forceps. The available space between two adjacent tissue samples 70
can also be limited, causing mounting and removal challenges in
embodiments of apparatus 10 that employ two or more tissue samples
70 on tissue sample holder 30. The limited footprint on tissue
sample holder 30 can be further exacerbated as the number of tissue
samples 70 mounted on tissue sample holder 30 are increased for a
given size blade 51 (FIG. 4), to increase the throughput of
apparatus 10.
[0050] With continued reference to FIG. 5, to reduce the likelihood
of the aforementioned difficulties in mounting and/or removing
tissue sample(s) 70 to and from tissue sample holder 30, some
embodiments of apparatus 10 can include one or more optional
intermediary structures, such as removable plug(s) 80 that are
removable from tissue sample holder 30 (e.g., from base 31).
Plug(s) 80 can comprise similar or different material as tissue
sample holder 30. In some embodiments, plug(s) 80 can comprise a
material with different adherence and/or structural properties than
tissue sample holder 30.
[0051] Plug(s) 80 can facilitate deployment and removal of a tissue
sample 70 to and from tissue sample holder 30. For example, tissue
sample(s) 70 can be removed from and/or mounted to plug(s) 80 while
plug(s) 80 are detached and separated from tissue sample holder 30,
to provide additional space during the removal/mounting procedure.
Plug(s) 80 can also facilitate deployment and removal of tissue
sample(s) 70 to and from tissue sample holder 30 without removing
tissue sample holder 30 from container 20.
[0052] Plug 80 can comprise any of a number of shapes, such as
those cross-sectional shapes described generally herein for tissue
sample holder 30, or other shapes. In some embodiments, plug 80 is
sized and shaped to support a single tissue sample 70, but it will
be understood that plug 80 can be sized and shaped to support two
or more tissue samples 70.
[0053] Referring to FIG. 5, plug(s) 80 can attach to tissue sample
holder 30 using any of a number of engagement mechanisms, such as
those described herein for attaching tissue sample holder 30 to
container 20, or other mechanisms. In the illustrative embodiment,
plug 80 includes a body 81 configured to engage with (e.g., slide,
or insert into) a corresponding structure (e.g., a slot 34) on
tissue sample holder 30. Body 81 can include one or more portions
extending along and from various portions of its sides or surfaces,
to provide additional engagement with tissue sample holder 30
and/or to provide an additional surface area on which a tissue
sample 70 can be mounted. For example, body 81 can include an upper
flange 82 extending from an upper portion of the sides of body 81,
to provide additional surface area for mounting a tissue sample 70,
and to provide further engagement with base 31 (e.g., upper surface
31a). For example, body 81 can include a lower flange 83 extending
from a lower portion of the sides of body 81 to provide further
engagement with a corresponding slot or other structure on base
31.
[0054] Plug(s) 80 can include one or more handling portions
configured to assist in the handling of plug(s) 80, including any
of those described herein for handling tissue sample holder 30, or
other alternatives. In the illustrated embodiment, plug(s) 80
include a handle or tab 84 extending from a portion (e.g., an
upper, lower or intermediary portion) of body 81.
[0055] Container 20 can comprise any of a number of shapes and
materials capable of receiving and containing a fluid, preferably a
liquid. For example, container 20 can comprise any of a number of
different cross-sectional shapes, such as an approximately
rectangular, elliptical, trapezoidal, or any other regular or
irregular shape that forms a hollowed, inner volume when extended
longitudinally, to hold fluid. Container 20 can comprise one or
more sidewalls 21 extending from a base 22 of container 20, to form
an internal volume within container 20. The positioning of base 22
is not limited to a lowermost extremity of container 20; base 22
can be positioned anywhere within an inner perimeter of sidewall(s)
21 to form an internal volume within container 20. An upper portion
of container 20 can be closed, e.g., with a cover or lid.
Preferably, a portion of container 20 (e.g., an upper or side
portion) is open, or is configured with one or more openings, to
facilitate insertion and removal of the tissue sample holder 30 to
and from container 20, and to allow a portion of microtome 50 to
section a portion 71 of tissue sample 70 supported on or within
container 20.
[0056] Container 20 can comprise any of the materials described
herein for tissue sample holder 30, and can comprise a similar or
different material. Container 20 preferably comprises a hydrophobic
(e.g., waterproof) material, such that container 20 can hold a
liquid, and more preferably, a material suitable for use within a
clinical environment for handling tissue samples. In one
embodiment, container 20 comprises polycarbonate or a similar
plastic. Container 20 can comprise an opaque, translucent, or
transparent material. It will be understood that container 20 can
comprise any combination of, and/or can be coated with, one or more
of the aforementioned materials.
[0057] Apparatus 10 can include one or more inlets 43 configured to
control (e.g., restrict and/or allow) fluid flow from a fluid
source into container 20. Inlet 43 can be separately or integrally
formed with respect to container 20. Inlet 43 can be attached to
container 20 or an intervening structure, or can be the top opening
of container 20, such that fluid can pour or flow from a position
separate from container 20 (e.g., above container 20) and into
container 20. Inlet 43 can comprise an opening extending through a
portion of container 20, such as one or more of sidewalls 21 and/or
base 22. In some embodiments, inlet 43 can include, or can be in
communication with, one or more of a fitting, spout, nozzle,
conduit, valve, flow controller, pump, pressure regulator or
similar fluid control device configured to control the delivery of
fluid into container 20. In FIG. 4, the fluid connection to inlet
43 is not shown for clarity. It will be understood that any such
fluid control devices known or described herein can be manually or
automatically controlled.
[0058] Apparatus 10 can include one or more drains configured to
control (e.g., restrict and/or allow) fluid flow from container 20.
Referring to FIGS. 4 and 5, apparatus 10 can include a level
control drain 27 configured to maintain a level of fluid within
container 20, such as a fluid level with respect to tissue sample
70 in container 20. In some embodiments, level control drain 27 can
be configured such that fluid can flow across substantially the
entirety of the height of tissue sample 70 when tissue sample 70 is
in container 20 and throughout the sectioning of tissue sample 70
by microtome 50. In some embodiments, level control drain 27 can be
configured such that the entirety of tissue sample 70 is immersed
in fluid, to reduce damage or contamination to a portion 71 of
tissue sample 70 sectioned by microtome 70. In some embodiments,
drain 27 can be configured to provide a given level of fluid in
container 20 that is a desired height above a tissue sample 70
mounted on tissue sample holder 30 in container 20. In some
embodiments, the height or position of drain 27 can be adjustable
with respect to a portion of container 20, tissue sample holder 30,
tissue sample 70, and/or outlet(s) 40. Level control drain 27 can
be separately or integrally formed with respect to container 20.
Drain 27 can include, or can be in communication with, any of the
fluid control devices known or described herein, configured to
control the drain of fluid from container 20. In some embodiments,
drain 27 can be a gravity drain. The connection of the drain 27 to
a fluid sink is omitted from FIG. 4 for clarity.
[0059] Referring to FIG. 5, in some embodiments, drain 27 can
comprise a drain opening or outlet 27a extending through a portion
of container 20 (e.g., any of sidewall(s) 21 and/or base 22), to
facilitate fluid flow from container 20. In some embodiments, drain
27 can comprise a drain inlet or opening 27b in fluid communication
with drain outlet 27a. Drain outlet opening 27a and inlet opening
27b can be in fluid communication with each other through a number
of different structures extending between outlet 27a and inlet 27b,
such as a tube, conduit, etc. In the illustrated embodiment, drain
27 can include a drain body 27c with a channel extending
therethrough that fluidly connects outlet 27a and inlet 27b.
[0060] Drain inlet 27b can be positioned at a given height with
respect to base 22 of container 20, to maintain the aforementioned
level of fluid within container 20. For example, drain inlet 27b
can be positioned at a height sufficiently above the upper portion
71 of tissue sample 70, when tissue sample holder 30 is mounted
within container 20, such that fluid can flow across substantially
the entirety of the height of tissue sample 70 throughout the
sectioning of tissue sample 70 by microtome 50. In some
embodiments, inlet 27b can be positioned such that the
cross-section of its opening is approximately parallel with the
surface of a fluid held within container 20, to maintain a
consistent fluid height within container 20 as the flow of fluid
within container 20 varies. In the illustrative embodiment, inlet
opening 27b extends through an upwardly-facing surface of drain
body 27c, which is positioned with respect to container 20 to be
approximately parallel with a fluid held within container 20.
[0061] The one or more inlet(s) 43, drain(s) 27, and/or outlet(s)
40 can be configured in any of a number of ways to facilitate
movement of a sectioned portion 71 of tissue sample 70 into outlet
40 after being sectioned by microtome 50. For example, the flow
rate of fluid through inlet(s) 43 can be adjusted to comprise a
smooth, laminar flow of fluid within container 20 towards outlet
40, without causing turbulence that might cause the sectioned
portion 71 of tissue sample 70 to move away from outlet 40. In some
embodiments, inlet(s) 43 can include and/or be in fluid
communication with a fluid control device that can be used to
adjust such flow rate. In some embodiments, a valve can be in
communication with inlet 43, to control the flow of fluid into
container 20; in such embodiments, said valve can be closed during
or subsequent to the sectioning of a portion 71 of tissue sample
70, to reduce fluid movement within container 20 and increase the
likelihood of the sectioned portion 71 of tissue sample 70 to move
into outlet 40 after the cutting stroke of microtome 50 is
completed. In these embodiments, there will be no fluid entering
the container 20 during the sectioning process, but fluid will be
flowing out of outlets 40 as described above. The fluid level will
drop during this time, but enough fluid can be present in the
container to maintain everything appropriately submerged. After the
sections exit the container 20 through outlets 40, the inlet can be
turned back on, and the container fluid is replenished for the next
cycle.
[0062] As described above, the flowpath of fluid within container
20 with respect to outlet(s) 40 and/or tissue sample(s) 70
facilitates the movement of a sectioned portion 71 of tissue sample
70 towards and into outlet 40. For example, inlet(s) 43 can be
aligned with or can point towards tissue samples 70 and outlet 40,
to direct fluid flow over tissue samples 70 and towards outlet 40.
In some embodiments, inlet 43 and/or a portion of container 20 can
include a channel 45 with a curvilinear path that facilitates a
smooth flow of fluid within container 20, and reduces the
likelihood of turbulence. In some embodiments, inlet 43 can be
positioned within container 20 and with respect to drain 27, such
that outlet(s) 40 and/or tissue sample holder 30 are positioned
between inlet 43 and drain 27. Such embodiments can provide a
flowpath between inlet 43 and drain 27 that allows fluid to flow
over tissue samples 70 on tissue sample holder 30, and facilitates
the flow of sectioned portions of tissue samples 70 towards
outlet(s) 40. For example, inlet 43 can extend through a first
portion (e.g., end) of a sidewall 21, with drain 27 extending
through a second portion (e.g., an opposed end) of the sidewall 21,
with tissue outlet(s) 40 extending through an intermediary portion
of the sidewall 21.
[0063] In some embodiments, apparatus 10 can include an overflow
drain 26 configured to maintain a level of fluid within container
20, for example, to prevent overflow of fluid from container 20.
Overflow drain 26 can be substantially similar to the
aforementioned embodiments of level control drain 27. Preferably,
overflow drain 26 can be configured with an inlet 26a that is
positioned with respect to container 20 and a fluid held within
container 20, to maintain a level of the fluid within container 20
that is greater than the level of fluid maintained by level control
drain 27. As such, inlet 26a can be a greater height than inlet 27b
of level control drain 27 with respect to a fluid surface within
container 20. Such an embodiment will prevent overflow of fluid
from container 20 (e.g., over the upper edges of sidewall(s) 21) in
the event of failure of drain 27 and/or in the event that the net
flow of fluid into container 20 (e.g., from inlet 43 or other
inlets) otherwise exceeds the flow of fluid from container 20 (e.g.
from drain 27 and/or other drains or outlets, such as outlets
40).
[0064] Continuing to refer to FIGS. 4-6, outlet(s) 40 can be
configured to allow flow of a fluid from the container 20, wherein
the flow of fluid through the outlet(s) 40 causes a portion of the
tissue sample sectioned by a microtome to move into the outlet(s)
40. Such movement can be facilitated through a pipette, a gravity
drain, a siphon, or a pump or similar fluid control device in
communication with outlet(s) 40. In embodiments with two or more
outlet(s) 40, a flow through a first outlet 40 can cause a portion
of a first tissue sample 70 to flow into the first outlet 40, and a
flow through a second outlet 40 can cause a portion of a second
tissue sample 70 to flow into the second outlet 40. It will be
understood that apparatus 10 can include any of a number of
different outlet(s) 40, and is shown with four outlets 40 for
illustrative purposes only. Additionally, outlets 40 are shown to
be approximately co-linear and co-planar, in a single row, although
some embodiments can include a plurality of non-linear or
non-planar outlets, and/or can form one or more rows and/or columns
of outlets. Advantageously, the number and orientation of the
outlets 40 is related to the number and orientation of the tissue
samples 70, as described further herein.
[0065] Outlet 40 can be separately or integrally formed with
respect to container 20. Outlet 40 can comprise an opening
extending through a portion of container 20, such as base 22, a top
or cover on container 20, or preferably, through a portion of
sidewall 21, to allow a portion 71 of tissue sample 70 to flow from
container 20. Thus, outlet 40 can be configured such that a
sectioned portion of a tissue sample 70 can flow vertically (e.g.,
upwardly or downwardly), or preferably, horizontally from container
20, or other angles with respect to container 20.
[0066] In some embodiments, the shape and/or size of outlet 40 can
be configured to facilitate the movement of a portion of a tissue
sample 70 into outlet 40. The cross-sectional shape of outlet 40
can be any of a number of different shapes, such as a substantially
elliptical, circular, rectangular, square, arc-shaped, round,
non-round, or other regular or irregular shape. Outlet 40 can be a
similar or different cross-sectional shape than tissue sample 70
and/or a sectioned portion 71 of tissue sample 70. In a preferred
embodiment, outlet 40 can comprise a cross-sectional shape that is
roughly similar to the side cross-sectional shape of an
un-sectioned tissue sample 70 (e.g., an elongated shape, such as a
rectangular or elliptical cross-section). Such similarity in shape
can facilitate movement of any portion 71 of tissue sample 70
towards outlet 40, regardless of which portion (e.g., an upper,
lower, or intermediary portion) of tissue sample 70 is being
sectioned. One or more dimensions of outlet 40 (e.g., height,
width, diameter) can be sized to be equal to or slightly larger
than a sectioned portion 71 of tissue sample 70, to avoid snagging
or interference when such sectioned portion moves into outlet
40.
[0067] In some embodiments, outlet 40 can include, or can be in
communication with, one or more fittings, spouts, nozzles,
conduits, valves, pumps, sensors, flow controllers, pressure
regulators or similar fluid flow devices configured to control the
flow of fluid through outlet 40 and the movement of sectioned
portions of tissue samples towards outlet 40. In some embodiments,
the amount of fluid flowed through outlet(s) 40 can be controlled
to one or more flow rates, to facilitate such movement. As
described above, and referring also to FIG. 2, in some embodiments,
the amount of fluid flowing through outlet(s) 40 can be controlled
to a first flow rate (e.g., a lower flow rate, or bleed) prior to
completed sectioning of a portion 71 of tissue sample 70 (e.g.,
before or during the sectioning of tissue sample 70, but prior to
completing a cutting stroke of a section of tissue sample 70 by
microtome 50), and a second (e.g., greater) flow rate after
completed sectioning of a portion 71 of tissue sample 70 (e.g.,
subsequent to completing a cutting stroke of microtome 50.
Controlling the fluid flowing through outlet 40 to a first lower
flow rate prior to completing the cutting stroke can help draw a
portion 71 of tissue sample 70 towards outlet 40 during the
sectioning cut, without causing the portion 71 to curl, roll, or
flap over onto itself, reducing the accuracy of the cut.
Controlling the fluid flowing through outlet 40 to a first lower
flow rate can also reduce the amount of time that the second,
increased flow rate need be applied to facilitate movement of the
sectioned tissue sample 70 to outlet 40, reducing fluid consumption
of apparatus 10
[0068] Referring to FIGS. 4 and 6, apparatus 10 can include one or
more conduit(s) 41, 42 in fluid communication with one or more
outlet(s) 40, to control the flow of fluid from container 20
through outlet(s) 40. For example, conduit 42 can be configured to
provide the first lesser bleed flow rate through outlet 40, and
conduit 41 can be configured to provide the second, greater flow
rate. Conduit 41 can facilitate flow from outlet 40 to storage
plate 90.
[0069] Conduits 41, 42 can comprise any of the cross-sectional
shapes and/or sizes described herein for outlets 40, and can
comprise a similar or different shape and/or size. In some
embodiments, conduits 41 and/or 42 can be tapered in size and/or
shape along their longitudinal length. For example, one or more
dimensions (e.g., height, width, and/or diameter) of the interior
cross-section of conduit section 41 can decrease in a longitudinal
direction with respect to outlet 40, to facilitate movement of the
sectioned portion 71 of tissue sample 70 from the outlet 40 through
conduit 41. In a preferred embodiment, the interior cross-sectional
shape of conduit 41 is tapered from an approximately elliptical
cross-sectional shape at its interface with outlet 40 to an
approximately circular cross-sectional shape downstream from outlet
40. In another preferred embodiment, conduit 41 comprises an inner
diameter of approximately 0.25 in.
[0070] Conduits 41, 42 can comprise any of a variety of rigid,
semi-rigid, or flexible hoses, channels, manifolds, or other
structures, comprising any of a variety of materials, known or
described herein suitable for channeling fluid. It will be
understood that conduits 41, 42 need not include a completely
enclosed lumen, and may comprise a partially open channel or
similar configuration suitable for channeling fluid. It will also
be understood that conduits 41 and 42 are optional, and that
apparatus 10 can be configured such that fluid and a sectioned
portion 71 of tissue sample 70 can flow (e.g., "pour,") directly
from outlet(s) 40.
[0071] Conduit 41 can comprise one or more optional conduit
sections. For example, conduit 41 can comprise a section 41a
extending from opening 40 and through a portion of sidewall 21.
However, it will be understood that section 41a can extend from
another portion of container 20, and need not extend through
sidewall 20 or be integrally formed therewith. In embodiments with
two or more openings 40, two or more sections 41a can be configured
to extend or separate outwardly (e.g., laterally) with respect to
each other along their longitudinal length, to provide increased
spacing between the downstream ends of adjacent sections 41a with
respect to the upstream ends (e.g., at the interface of conduit(s)
41 with outlet(s) 40) (FIG. 4).
[0072] Apparatus 10 can include an optional manifold 130 through
which a portion of conduit 41 (e.g., a section 41b) can extend.
Section 41b can be integrally or separately formed with respect to
manifold 130. Section 41b can be connected to and in fluid
communication with conduit section 41a. Manifold 130 can include
one or more fittings 131 connected to a manifold body 132,
configured to connect and fluidly communicate with opening(s) 40,
inlet 43, and/or drain 27. Fittings 131 can be a push-connect,
barb, compression style, or other type of fluid connector. In some
embodiments, manifold 130 can comprise a quick-connect, luer
connection, or other removable connection 133 that facilitates the
removal and reattachment of manifold 130 to opening(s) 40, inlet
43, and/or drain 27.
[0073] Conduit 41 can comprise a channel or conduit section 41c
connected to and in fluid communication with section 41b. Conduit
section 41c can be connected to section 41b, for example, with
fitting 131. Section 41c can provide movement of fluid and a
sectioned portion 71 of tissue sample 70 from outlet 40 to another
device, such as storage plates 90.
[0074] Apparatus 10 can include one or more valves 120 in fluid
communication with one or more outlet(s) 40, and configured to
control the flow of fluid from the outlet 40. Valve 120 can be in
communication with and mounted directly to outlet 40 (e.g., a
portion of sidewall 21), or on an intermediary structure in
communication with outlet 40, such as manifold 130 and/or a section
of conduit 41. Valve 120 can comprise any of a number of different
types of valves, such as a ball valve, gate valve, pinch valve,
spool valve, diaphragm valve, butterfly valve, etc. Valve 120 can
be actuated manually, hydraulically, electrically (e.g., a solenoid
valve), and/or pneumatically. Preferably, valve 120 can comprise a
valve with a configuration and/or materials suitable for use in a
clinical environment, to reduce the likelihood of contamination of
sectioned portions of tissue sample(s) 70.
[0075] Valve 120 can comprise a bleed, or non-fully closed valve
that allows a greater flow in its open position, and a lesser flow
in its closed position, or can be a shutoff valve that restricts
any significant flow in a closed position. In one embodiment, valve
120 can comprise a pinch valve configured to control (e.g.,
restrict or allow) flow through a portion of conduit 41. In the
illustrated embodiment, valve 120 controls flow from conduit
section 41c and through an outlet conduit section 41d downstream of
valve 120. Valve 120 can control flow from outlet 40 by clamping
(e.g. pinching) and unclamping conduit 41 between an actuator,
cylinder, or other clamping means 121 and a mandrel or other
clamping means 122. Using a pinch valve configuration can clamp or
pinch conduit 41 between an open and closed position, to allow and
restrict flow, respectively, with reduced likelihood of contact
between the internal components of valve 120 and the fluid or
sectioned portions 71 of tissue sample 70 flowing through valve
120. As used herein, the "closed" position of a valve can allow a
bleed or a lesser flow therethrough, for example, when valve 120
comprises a non-fully closed valve. A pinch valve can reduce the
likelihood of contamination or damage to the sectioned portions 71
of tissue sample 70.
[0076] Valve 120 can be closed to provide a lower flow rate through
outlet 40 prior to completing the cutting stroke during the
sectioning of the portion 71 of tissue sample 70. When valve 120 is
closed, the lower flow rate through outlet 40 can be approximately
zero (for example, if valve 120 is a shutoff valve), or can
comprise a flow greater than zero through outlet 40 (for example,
if valve 120 is a non-fully closing or bleed valve, or in
embodiments that allow flow through optional conduit 42, described
further herein). Valve 120 can be opened to provide an increased
flow rate subsequent to sectioning a portion 71 of tissue sample
70, to allow an increased fluid flow towards outlet 40, moving the
sectioned portion of the tissue sample towards and into outlet 40.
The amount of flow through outlet 40 when valve 120 is opened can
be varied; in a preferred embodiment, the flow through outlet 40
when valve 120 is opened is approximately 500 mL/min.
[0077] Optional conduit 42 can be included with apparatus 10 to
facilitate movement of a sectioned portion 71 of tissue sample 70
into outlet 40. Conduit 42 can be configured to flow an amount of
fluid through outlet 40 that is advantageously less than the amount
of fluid flowed through outlet 40 by conduit 41. For example,
conduit 42 can be configured to provide the aforementioned lower
flow rate, or bleed, through outlet 40 prior to completing the
cutting stroke during the sectioning of the portion 71 of tissue
sample 70. Conduit 42 can provide lesser fluid flow through outlet
40, for example, by sizing the inner channel of conduit 42 to be a
smaller cross-sectional area than that of conduit 41. Additionally
or alternatively, conduit 41 and/or 42 can include any fluid
control devices known or described herein that are configured to
provide different flow rates than the other of conduit 41 and 42,
or flow devices in fluid communication therewith. For example, an
optional flow controller or needle valve 44 can be in communication
with conduit 42 to control the flow therethrough. In an embodiment,
needle valve 44 and/or conduit 42 can be configured to allow a
bleed flow rate of approximately 25 ml/min.
[0078] It will be understood that the lower flow provided by
conduit 42 can be implemented as an alternative or in combination
with a lower flow (e.g., bleed) provided with conduit 41 (e.g.,
when valve 120 comprises a bleed valve). It will also be understood
that the bleed through conduit 42 can be continuous (e.g., used
when conduit 41 provides a higher flow rate), or can be switched
sequentially with respect to a higher flow rate through conduit 41
(e.g., when valve 120 is opened).
[0079] In the illustrated embodiment, conduit 42 can be in fluid
communication with outlet 40 at a connection point 44 located in
first conduit 41. Connection point 44 can comprise any of a number
of structures, such as a tee-fitting, y-fitting, manifold, banjo
fitting, and the like. In some embodiments, connection point 44 can
comprise a valve that controls (e.g., restricts or allows) fluid
communication between outlet 40 (e.g., portion 41c of conduit 41),
and either or both of conduit 42 and conduit section 41(d). The
fluid control devices known or described herein can be positioned
to control flow in conduits 41 and/or 42 upstream and/or downstream
of connection point 44. In some embodiments, a filter can be
positioned within conduit 41 and/or 42 to allow the flow of fluid
into conduit 42, but prevent the likelihood of a sectioned portion
71 of tissue sample 70 flowing into conduit 42.
[0080] Referring to FIGS. 4 and 7, a storage plate 90 can be
configured to include one or more wells 91 configured to receive
one or more sectioned portion(s) of tissue sample(s) 70. For
example, after a portion 71 of tissue sample 70 is sectioned by
microtome 50, it can flow through outlet 40, and from outlet 40
into well 91. In some embodiments, the portion 71 of tissue sample
70 flows into well 91 from an end of conduit 41, such as section
41c. Storage plate 90 and/or well(s) 91 can comprise any of the
shapes and/or materials described generally herein for container
20.
[0081] Storage plate 90 and well(s) 91 can comprise any of a number
of suitable configurations to receive a sectioned tissue sample. In
the illustrated embodiment, storage plate 90 comprises one or more
sidewalls 93 extending from a base 95 to form an interior volume
configured to receive one or more sectioned tissue samples 70.
Preferably, storage plate 90 comprises one or more inner dividing
walls 94 to form a plurality of wells 91. One or more apertures 92
can extend through a portion of base 95, to allow fluid to flow
through a portion of the well without allowing a tissue sample
section to flow through the aperture(s) 92. Apertures 92 can also
increase the storage density of the sectioned portions of tissue
samples 70 that can be stored in wells 91, by reducing the amount
of fluid within wells 91, and allowing the sectioned portions to
stack, contact each other, and/or otherwise be stored in a more
efficient, compact manner. It will be understood that the number of
wells 91 in each storage plate 90 (e.g., the illustrated quantity
of 24, in a 4.times.6 matrix), and/or the number of apertures 92 in
each well can be any of a number of quantities, and the quantities
shown in FIGS. 4 and 7 are for illustrative purposes only.
[0082] Referring to FIGS. 4, 7 and 8, apparatus 10 can include tray
110 configured to support the one or more storage plates 90. Tray
110 can comprise any of the shapes and/or materials described
herein generally for container 20. Tray 110 can comprise a base 112
with one or more sidewalls 111 configured to form an interior
volume in which storage plate(s) 90 can be contained and supported.
Tray 110 can include one or more optional supporting members 113
(e.g., brackets) attached, e.g., to sidewall(s) 111 and/or base
112, to provide additional support to the one or more storage
plate(s) 90. Such additional support can improve the accuracy of
the transfer of sectioned tissue samples into storage plate(s) 90.
Tray 110 can be sized to allow storage plate(s) 90 to remain
submerged under fluid, to protect tissue sample sections stored
within storage plate(s) 90. Tray 110 can comprise one or more
drains to control the flow of fluid from tray 110, similar to
drains 26, 27 of container 20 (FIGS. 4-5), and to manage the flow
of fluid through aperture 92 in storage plate 90, and into tray
110.
[0083] Tray 110 can be moveable with respect to one or more other
portions of apparatus 10, such as microtome 50, container 20,
outlet(s) 40, and/or conduit(s) 41 (e.g., the conduit outlet of
section 41(d)). Such movement can allow tray 110 (and thus storage
plates 90) to be indexed with respect to outlet(s) 40. This
indexing can allow a first sectioned portion of a tissue sample 70
to be flowed from outlet 40 into a first well 91 on a storage plate
90, after which storage plate 90 can be moved (e.g., in one or more
of directions 501, 502, 505, 506), such that storage plate 90 can
subsequently receive a second sectioned portion of a tissue sample
70 from outlet 40 into first well 91 or a second well 91. The
relative movement between tray 110 and another portion of apparatus
10 can be provided manually or automatically through any of a
number of movable elements, such as those known or described herein
for providing relative movement between microtome 50 and tissue
sample(s) 70.
[0084] In some embodiments, apparatus 10 can include one or more
optional sensors (e.g., pressure, temperature, proximity, location
(e.g., an encoder), flow, fluid level sensors, etc.) to provide
feedback, and/or to allow for open or closed loop control, or
manual adjustment, of various aspects of apparatus 10. For example,
an optional sensor 46 (FIG. 6) can be employed proximate to or
within outlet 40 and/or conduit 41, to detect when (and if) a
sectioned tissue sample successfully passes through outlet 40
and/or conduit 41.
[0085] In some embodiments, apparatus 10 can include a temperature
control system to control the temperature of (e.g., heat or cool)
various components of apparatus 10, or materials being worked upon
by apparatus 10. For example, it may be advantageous to control the
temperature of the fluid flowing through or across one or more of
container 20, tissue sample holder 30, outlet(s) 40, drain(s) 26,
27, conduits 41, 42, storage plate 90, tray 110, and/or other
components described herein. Such temperature control can maintain
the integrity of a tissue sample or section as it is supported,
sectioned, moved, and/or stored by apparatus 10. Such a temperature
control system can include one or more heating and/or cooling
elements, and/or temperature sensors to control said heating and/or
cooling elements in a closed or open-loop control configuration. In
some embodiments, the temperature of the fluid in a reservoir that
feeds the container is maintained at a desired low temperature. The
fluid may be cycled through the system at a rate that significant
temperature increase when the fluid is in the system is avoided and
the only cooling element is associated with the reservoir rather
than with any of the fluid handling components described in detail
herein.
[0086] Referring again to FIGS. 2-4, in some embodiments, the
method of handling can further comprise flowing a fluid from the
one or more outlets 40 to storage plate 90, causing the one or more
portions of the tissue sample 70 to move from the one or more
outlets 40 into one or more wells 91 of the storage plate. In some
embodiments, the method can further comprise repeatedly flowing a
fluid from the at least one outlet 40 to storage plate 90, and
indexing the storage plate 90 from a first position to a second
position between steps of flowing a fluid from the one or more
outlets 40 to the storage plate 90, causing a first portion 71 of
the tissue sample 70 to move into a first well 91 during the first
step, and a second portion of the tissue sample to move into a
second well 91 during the second step. In some embodiments, flowing
the fluid from the one or more outlets 40 to a storage plate can
comprise flowing the fluid from a plurality of outlets 40 to a
plurality of storage plates 90.
[0087] The container 20, tissue sample holder 30, plug 80, storage
plate 90, tray 110, and/or microtome 50 described herein may be
attached together at the point of manufacture. Alternatively, any
of these components or others described herein can be manufactured
as an accessory or replacement part and sold independently. For
example, container 20 can be sold without a tissue sample holder
30; tissues sample tissue sample holder 30 can be sold without plug
80; tray 110 can be sold without storage plate 90, etc. Any
combination of container 20, tissue sample holder 30, plug 80,
storage plate 90, tray 110, and/or microtome 50 can also be
supplied as a kit, for example, wherein the container 20, tissue
sample holder 30, plug 80, storage plate 90, tray 110, and/or
microtome 50 can be supplied separately and then assembled by the
user.
[0088] Embodiments described herein provide less expensive,
efficient apparatuses and processes for handling a portion of a
tissue sample sectioned by a microtome that can provide increased
quality, accurately sectioned tissue samples, with reduced waste
and contamination. Some embodiments provide a level of
repeatability and accuracy that can allow 120-140 portions of
tissue samples to be sectioned from a single mouse brain. Some
embodiments allow one or more sectioned portions of one or more
tissue samples to remain immersed in fluid throughout one or more
operations, and preferably, the entirety, of the sectioning,
handling, and storage process, to prevent contamination or damage
to the sectioned tissue samples. Some embodiments allow one or more
portions of a tissue sample to be sectioned, handled, and/or
stored, without requiring contact by laboratory personnel or
external tools (e.g., forceps). Some embodiments allow one or more
of the sectioning, handling and/or storage steps to be controlled
automatically, reducing or eliminating manual input from laboratory
personnel during these step(s). Some embodiments allow two or more
portions of two or more tissue samples to be sectioned during the
same cutting stroke of a microtome blade.
[0089] The above-described embodiments have been provided by way of
example, and the present invention is not limited to these
examples. Multiple variations and modifications to the disclosed
embodiments will occur, to the extent not mutually exclusive, to
those skilled in the art upon consideration of the foregoing
description. Additionally, other combinations, omissions,
substitutions and modifications will be apparent to the skilled
artisan in view of the disclosure herein. Accordingly, the present
invention is not intended to be limited by the disclosed
embodiments.
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