U.S. patent number 3,771,490 [Application Number 05/228,115] was granted by the patent office on 1973-11-13 for automatic tissue processor.
Invention is credited to Thomas D. Kinney, John E. P. Pickett.
United States Patent |
3,771,490 |
Kinney , et al. |
November 13, 1973 |
**Please see images for:
( Certificate of Correction ) ** |
AUTOMATIC TISSUE PROCESSOR
Abstract
A tissue processor is programmed for fixation, dehydration and
clearing of electron microscopy size particles and utilizes a
porous receptacle for each particle or group of particles of tissue
and a processing chamber adapted to contain a plurality of the
receptacles. The chamber is connected to a plurality of containers,
some of which are refrigerated, and which contain the various
processing solutions. The solutions are individually piped to, and
from beneath, the chamber through a remotely controlled valve and
manifold arrangement which also connects with a metering pump. This
arrangement minimizes fluid contamination and allows each solution
to be precisely metered, brought to, retained in and drained from
the chamber thus allowing the particles to be bathed in the
solutions according to an automatic programmed time sequence. The
program may be varied as to number of cycles per solution, as to
number of solutions per program, as to time per cycle and as to
starting and terminal solutions in the program.
Inventors: |
Kinney; Thomas D. (Durham,
NC), Pickett; John E. P. (Durham, NC) |
Family
ID: |
22855857 |
Appl.
No.: |
05/228,115 |
Filed: |
February 22, 1972 |
Current U.S.
Class: |
118/698; 118/702;
118/421; 222/136 |
Current CPC
Class: |
G01N
1/31 (20130101) |
Current International
Class: |
G01N
1/31 (20060101); G01N 1/30 (20060101); B05c
011/10 () |
Field of
Search: |
;118/4,7,8,421,429,50
;134/96,97,98,195 ;137/1NE ;222/136,144.5,144,190 ;95/93 |
References Cited
[Referenced By]
U.S. Patent Documents
Primary Examiner: Kaplan; Morris
Claims
I claim:
1. A closed processing system enabling a plurality of individual
particles of minute electron microscopy size, having at least one
dimension greater than 1 millimeter, to be separately contained
while being bathed simultaneously for varying lengths of time in
successive selected aqueous and non-aqueous electron microscopy
tissue solutions so as to fix, dehydrate and clear the particles
preparatory to embedding, comprising:
a. a pluraltiy of uniform porous tissue receptacles each being
adapted to contain and physically isolate a group of said particles
having at least one particle per group, each receptacle having a
body portion defining an open cavity adapted to receive and retain
a group of particles and closure means removably received by said
body to enclose said cavity, the material forming said receptacles
being inert to all of said solutions and having for each of said
solutions at least some porous portion permeable thereto;
b. cabinet means providing a housing and respective refrigerated
and non-refrigerated compartments therein;
c. a plurality of closed solution containers selectively mounted in
said refrigerated and non-refrigerated compartments and each
containing a particular electron microscopy tissue processing
solution and collectively containing all of said aqueous and
non-aqueous solutions;
d. a discharge pipe and an associated remotely and electrically
controllable valve for each container connected to provide an
individual selectively controllable flow path for each of said
solutions to one side of its respective corresponding valve,
selected said pipes having at least a portion thereof positioned so
as to be cooled in said refrigerated compartment;
e. a manifold structure fixedly mounted in said housing adjacent
said compartments and mounting said container valves, said manifold
structure providing about the periphery thereof an inlet port for
and communicating with an opposite side of each respective said
container valve, a centrally disposed, vertically directed
passageway extending through said manifold, each said inlet port
being communicated to said vertically directed passageway by a
serparate one of a plurality of radially extending passageways, a
pump port located in said periphery and a further radial passageway
communicating said pump port with the vertical passageway;
f. a processing chamber mounted in said housing adjacent and above
said manifold structure, said chamber having a removable top cover
and being adapted to receive and process selected numbers of said
receptacles simultaneously, said chamber having an entry-discharge
port at the bottom thereof and a remotely and electrically
controllable valve located between and controlling communication
between said entry-discharge port and the top end of said vertical
passageway;
g. remotely controllable metering pump means and positioned within
said housing adjacent said manifold structure and having a pump
inlet communicating with said manifold structure pump port whereby
said pump is enabled to withdraw and discharge through said pump
port a positively measured volume of a selected said solution;
h. waste solution collection means including a waste discharge
remotely electrically controllable valve communicating on one side
with the bottom end of said vertical passageway and on an opposite
side with a waste solution collector whereby when said waste valve
is opened all solution fluids in said manifold passageways may flow
to said waste collector for disposal; and
i. remotely operable electrical control means for remotely
operating said container, chamber and waste valves and said pump
means in a predetermined time sequence program whereby with a
selected number of said receptacles installed in said chamber and
said cover on said chamber said solutions are successively,
selectively and independently drawn, externally measured, and
admitted to said chamber in a predetermined volume, retained for
predetermined times and drained from said chamber in a
corresponding processing sequence such that selected of said
aqueous and non-aqueous solutions are exchanged with each said
group of particles by penetration and drainage through said pores
while maintaining said system closed and each said group physically
isolated.
2. A system as claimed in claim 1 wherein said control means
include adjustable timing means enabling said predetermined time of
retention to be changed both with respect to all as well as
selected ones of said solutions.
3. A system as claimed in claim 1 wherein said control means
includes adjustable cycle switching means enabling selected
solutions to be cycled more than once through said chamber.
4. A system as claimed in claim 1 wherein said control means
includes time unit switching means enabling said predetermined time
of retention to be varied with respect to selected said
solutions.
5. A system as claimed in claim 1 wherein said control means
includes hold-pass switching means enabling selected said solutions
in one mode of switching to be by-passed and in another mode of
switching to constitute a program terminal stage.
6. A system as claimed in claim 1 wherein said control means
includes level selection switching means enabling a plural
selection of volumes pumped into said chamber by said pump
means.
7. A system as claimed in claim 1 wherein said control means
includes switching means enabling any said solution to be a
beginning stage in the said program.
8. A system as claimed in claim 1 wherein said control means
includes program switching means adapted upon actuation to select
certain said solutions and valves in a coordinated sequence adapted
to flush selected said manifold and chamber surfaces for cleaning
purposes.
9. A system as claimed in claim 1 wherein said manifold structure
comprises a vertically disposed substantially cylindrical block
containing said ports and passageways and having said container
valves and pump means mounted on the side thereof, said chamber
valve being mounted on an upper face and said waste valve on a
lower face thereof.
10. A system as claimed in claim 1 having fan exhaust and fume
collection means adapted to allow during operation of said system
continuous withdrawal of solution fumes both from within said
housing and from the area surrounding the top of said chamber.
11. A system as claimed in claim 1 wherein said control means
includes:
a. adjustable timing means enabling said predetermined times of
retention to be changed with respect to all said solutions,
b. adjustable cycle switching means enabling selected solutions to
be cycled more than once through said chamber,
c. time unit switching means enabling said predetermined time of
retention to be varied with respect to selected said solutions,
d. hold-pass switching means enabling selected said solutions in
one mode of switching to be by-passed and in another mode of
switching to constitute a program terminal stage,
e. level selection switching means enabling a plural selection of
volumes to be pumped into said chamber by said pump means,
f. switching means enabling any said solution to be a beginning
stage in a program, and
g. switching means adapted upon actuation to select certain said
solutions and valves in a coordinate subprogram sequence adapted to
flush selected said manifold and chamber surfaces for cleaning
purposes.
12. A system as claimed in claim 1 including means to vibrate said
receptacle during processing.
13. A system as claimed in claim 1 including means to agitate the
selected said solution in said chamber during processing.
14. A system as claimed in claim 1 wherein said chamber is mounted
in said housing immediately adjacent said refrigerated
compartment.
15. A system as claimed in claim 1 wherein said containers, pipes,
valves, manifold structure, chamber and pump means are stationary
within said housing.
16. A processing system for electron microscopy, light microscopy,
staining and similar tissue processing enabling a plurality of
individual tissue specimens to be bathed simultaneously for varying
lengths of time in successive tissue solutions preparatory to other
procedures, comprising:
a. holder means adapted to contain and physically isolate a
plurality of said specimens from each other, the material forming
said holder means being inert to all of said solutions and having
for each of said solutions at least some portion permeable
thereto;
b. cabinet means providing a housing and respective compartments
therein;
c. a plurality of closed solution containers mounted in said
housing and each containing a particular tissue processing
solution;
d. a discharge pipe and an associated remotely and electrically
controllable valve for each container connected to provide an
individual selectively controllable flow path for each of said
solutions to one side of its respective corresponding valve;
e. a manifold structure fixedly mounted in said housing adjacent
said compartments and mounting said container valves, said manifold
structure providing about the periphery thereof an inlet port for
and communicating with an opposite side of each respective said
container valve, a centrally disposed, vertically directed
passageway extending through said manifold, each said inlet port
being communicated to said vertically directed passageway by a
separate one of a plurality of radially extending passageways, a
pump port located in said periphery and a further radial passageway
communicating said pump port with the vertical passageway;
f. a processing chamber mounted in said housing adjacent and above
said manifold structure, said chamber having a removable top cover
and being adapted to receive said holder means and to process
selected numbers of said specimens simultaneously, said chamber
having an entry-discharge port at the bottom thereof and a remotely
and electrically controllable valve located between and controlling
communication between said entry-discharge port and the top end of
said vertical passageway;
g. remotely controllable metering pump means positioned within said
housing adjacent said manifold structure and having a pump inlet
communicating with said manifold structure pump port whereby said
pump is enabled to withdraw and discharge through said pump port a
positively measured volume of a selected said solution; and
h. remotely operable electrical control means for re-motely
operating said container and chamber valves and said pump means in
a predetermined time sequence program whereby with a selected
number of said specimens installed in said chamber and said cover
on said chamber said solutions are successively, selectively and
independently drawn, externally measured, and admitted to said
chamber in a predetrmined volume, retained for predetermined times
and drained from said chamber through valvular means connected to
said vertical passageway in a corresponding processing sequence
such that selected of said solutions are exchanged with each said
specimen while maintaining said system closed and each said
specimen physically isolated from the other.
17. A system as claimed in claim 16 wherein said manifold structure
comprises a vertically disposed substantially cylindrical block
containing said ports and passageways and having said container
valves and pump means mounted on the side thereof.
18. A system as claimed in claim 17 having fan exhaust and fume
collection means adapted to allow during operation of said system
continuous withdrawal of solution fumes both from within said
housing and from the area surrounding the top of said chamber.
Description
CROSS REFERENCE TO RELATED APPLICATIONS
This application is related to copending applications Ser. No.
860,256, filed Sept. 23, 1969, now U.S. Pat. No. 3,697,299 entitled
"Microscopy Tissue Receptacle Method" and Ser. No. 60,798, filed
Aug. 4, 1970, now abandoned entitled "Electron Microscopy Tissue
Processing Method."
BACKGROUND OF THE INVENTION
1. Field of the Invention
This invention relates broadly to apparatus and devices for
processing pieces of tissue but particularly to apparatus for
processing electron microscopy size tissue.
2. Description of the Prior Art
This invention constitutes an improvement upon the subject matter
of our prior U.S. Pat. No. 3,526,203, upon the subject copending
applications and upon the methods related thereto. Since the
general history of the prior art is dealt with in some detail in
prior U.S. Pat. No. 3,526,203 reference is made to U.S. Pat. No.
3,526,203 for the prior art background of the present invention.
Reference is also made to prior U.S. Pat. No. 3,536,040 directed to
a selectively permeable receptacle and to U.S. Pat. No. 3,576,176
directed to a tissue capsule rack which forms part of the prior art
of specific interest and useful in practicing the present
invention. In summary, the subject U.S. Pat. No. 3,526,203
discloses a processor for processing electron microscopy size
tissue, as well as larger tissue, in a permeable receptacle of the
type disclosed in U.S. Pat. No. 3,536,040 with the receptacle being
held in a rack as shown in U.S. Pat. No. 3,576,176. Our own prior
art is believed to be the closest known prior art. Of particular
interest to the present invention is that the processor of our
prior patent disclosed a system of connecting the processing
chamber with the processing solution containers by means of pipes
which connected to the top of the processing chamber and depended
on gravity feed and further depended on sensing the level of the
solution in the chamber as a means of stopping flow and measuring
the volume of a particular solution. After extensive experimental
use it was found that the processor of our prior patent inherently
introduced substantial opportunity for the solutions to mix in the
various pipes between the chamber and the containers. Also, after
experimental use with various types of solutions, particularly
osmic acid, and time programs the prior processor was found to
introduce a problem in maintaining accurate level control for the
various solutions. Certain solutions when passed through relatively
long lines tended to precipitate in the lines particularly when the
solution was held at or above room temperature and no provision was
made in the prior processor for maintaining selected solutions at
temperatures below room temperature.
Of particular interest is the fact that while our prior patent
taught the general concept of automatic operation with a program
switch the processor of the prior patent did not disclose means for
conveniently changing the numbers of cycles for a particular
solution. That is, in hand processing it is sometimes the practice
to hold the tissue in a solution, then drain the solution, then
hold the tissue again in the same solution for another time period.
The processor of our prior patent did not provide a means for
duplicating in a machine operation this aspect of hand processing.
While the prior patent processor contemplated changing an entire
program in order to obtain such a variation in numbers of cycles
per solution the processor of our prior patent did not not offer
the possiblity of making such a change even while a particular
program was in progress. Furthermore, the program capability of the
processor of our prior patent was to some extent limited in being
able to vary the amount of time devoted to a particular solution
being in the processing chamber. Thus, with the processor and
processing method of our prior patent if it was desired to change
the base time during which a particular solution was held in the
processing chamber from, say, 12 minutes to 24 minutes it was
necessary to change the entire program for the entire run of
solutions. Also, the prior processor did not exhibit the capability
of being able to pass certain solutions in a program or to
terminate a particular program, at least temporarily, with a
particular solution. In hand processing of electron microscopy
tissue it is sometimes desirable to, say, stain a tissue after
processing solutions 1, 2 and 3 and to, say, bypass solution 4 with
a certain tissue.
From the foregoing it can thus be seen that while the processor of
our prior patent constituted an important advance in the art and so
far as we know constituted the first significant step toward a
practical automatic processor for processing electron microscopy
size tissue long and extensive experimental use with that processor
have revealed a number of needed improvements in terms of both
apparatus and such constitutes the object of the present
invention.
SUMMARY OF THE INVENTION
The invention is generally directed to an apparatus for processing
electron microscopy size tissue and constitutes an improvement over
the apparatus shown called for in our prior U.S. Pat. No.
3,536,203. Reference to our prior patent is made for an
understanding of the general operation involved with an apparatus
of this kind. Of specific interest to the present invention, the
apparatus embodies a stationary processing chamber having a bottom
port connected directly to a manifold which is positioned below the
chamber. The manifold is also provided with a plurality of inlets
connected through pipes to each of the respective processing
solutions and with an outlet for waste discharge. Each inlet on the
manifold mounts a remotely solenoid controlled valve so that the
solutions from each solution container can be effectively isolated
from each other at points immediately adjacent the manifold which
minimizes the opportunity for, say, solution 1 to mix with solution
3 and also minimizes the amount of necessary waste solution. The
chamber port communicates with a solenoid control valve immediately
adjacent the manifold as does the waste outlet. A metering pump
also connects to the manifold and incorporates a metering piston
driven by a reversible motor so that the piston can be withdrawn to
pull in a given amount of solution and can be moved in the opposite
direction to force the solution into the processing chamber through
the mentioned bottom port. The manifold thus provides a passage for
a selected solution to be withdrawn into the pump and to then be
forced into the processing chamber. During withdrawal all valves
except the selected solution valve are closed. On the return stroke
only the processing chamber valve is opened and during dumping only
the waste valve is opened to allow the expended solution to drain
by gravity. Means are provided whereby the amount of pump cylinder
movement can be precisely regulated to vary the amount of solution
withdrawn so as to eliminate the need for level sensing and control
as in the processor of our prior patent. The various solenoid
valves are also coordinated by program switch means such that all
the respective solutions may be metered, pumped up to the
processing chamber and then allowed to drain by gravity to waste
with minimal mixing of one solution with another except in the
collective waste.
For greater variety of programming, the invention also adopts the
concept of establishing a so-called base time of, say, 15 minutes
and incorporating means whereby a particular solution may be held
during a particular cycle for one, two or three periods of base
time. Thus, merely by moving a switching arrangement a solution can
be held for, say, 15, 30 or 45 minutes. This can be further varied
so that, during a particular program, solution 1 stays for, say, 15
minutes, solution 2 for 45 minutes and solution 3 for 45 minutes in
that sequence. The program can be changed while in progress or from
program to program merely by reorienting a set of switching
positions. Thus, a medical research investigator is offered a wide
range of processing techniques for comparing one mode of processing
the same tissue or different tissue particules with another mode of
processing. The processor of the invention also mounts a switching
arrangement adapted to allow any particular solution to be bypassed
in a program or for the processing to stop, at least temporarily,
when such solution is reached. An improved ventilation system has
also been provided which allows fumes in the work area and
particularly around the processing chamber to be withdrawn as
rapidly as formed.
In summarizing the present invention in apparatus terms, it can be
said that the programming capability has been substantially
increased, the opportunity for solution mixing has been minimized
and control over the quantity of solution used has been made much
more precise. In method terms, it can also be said that the method
of the invention minimizes intermixing of solutions, increases
accuracy of measuring and provides greater variety in programming.
Other improvements of an important but lesser nature will also be
noted as the description proceeds.
DESCRIPTION OF THE DRAWINGS
FIG. 1 is a perspective view of an apparatus for processing
electron microscopy tissue according to the invention.
FIG. 2 is a right elevation view of the apparatus.
FIG. 3 is a rear elevation view of the apparatus.
FIG. 4 is a left elevation view of the apparatus with various
compartment doors open.
FIG. 5 is an enlarged front elevation view of the main control
panel and also showing the auxiliary control panel in an open
position.
FIG. 6 is a front elevation view of the solution containers with
the container compartment doors being shown in an open
position.
FIG. 7 is a perspective somewhat schematic view of the manifold,
metering pump, processing chamber and waste tank arrangement.
FIG. 8 is a sectional and somewhat schematic elevation view of the
chamber, valve and metering pump arrangement.
FIG. 9 is a plan, sectional and somewhat schematic view of the
manifold, passages, valves and metering pump.
FIG. 10 is a schematic diagram of the control system.
FIG. 11 is a perspective view of a prior art porous receptacle used
with the invention.
DESCRIPTION OF THE PREFERRED EMBODIMENT
The present invention is directed to processing apparatus and a
method for processing tissue particles held in a porous receptacle
25 of the type shown in FIG. 11 and more fully disclosed in related
U.S. Pat. No. 3,536,040. A typical receptacle of this type is
porous at least in some portion, whether in the body 20 or the end
caps 21, to all the solutions. It may be roughly one-half inch in
length and one-half inch in diameter and the pore size in the range
of about 40 to 200 microns. A substantial number of such
receptacles may be processed simultaneously and according to a time
schedule for the various processing solutions as later
discussed.
The processor of the invention includes a cabinet 29 having pivoted
front doors 30, 31, a pivoted rear door 32 and a pivoted top door
33. Doors 30, 31 cover, respectively, a refrigerated compartment 40
and a non-refrigerated compartment 41 having a dividing partition
42 and a back wall partition 43. Suitable heat insulation 45 is
mounted on door 31 and surrounding compartment 40 in order to
retain the reduced temperature obtained by suitable refrigeration
mechanism generally indicated at 46. Ten solution containers 50-59
are shown by way of example and are labeled respectively: solution
1, 4 percent glutaraldehyde; solution 2, buffer; solution 3, 1
percent osmium tetroxide; solution 4, 50 percent alcohol; solution
5, 70 percent alcohol; solution 6, 80 percent alcohol; solution 7,
95 percent alcohol; solution 8, absolute alcohol; solution 9,
propylene oxide; solution 10, propylene oxide-Epon 50-50. The first
three mentioned solutions have been found to require refrigeration
for best processing results and it is for this reason that
compartment 40 is refrigerated. Each solution container is provided
with a suitable quick-disconnect 70, FIG. 6, and an air inlet 71 to
allow the solution to be withdrawn through a connecting tube 72.
The respective connecting tubes 72 are each connected to a
respective solenoid valve 73 which in turn are mounted on a
manifold structure 75. Structure 75, as shown in FIG. 9, provides a
series of short passages for communicating all of the various
solution, chamber, waste and pump inlets and ports. Like other
components exposed to the solutions, structure 75 must be of a
material inert to all the solutions and stainless steel and Teflon
plastic have been employed. The ten tubes 72 leading from the ten
solution containers 50-59 are connected to ten respective solenoid
valves 73. Manifold 75 also mounts above the manifold a solenoid
valve 80 having a connecting pipe 81 connected to the processing
chamber 90 and a pipe 82 connected to the manifold 75. Below the
manifold there is mounted a further solenoid valve 85 having a pipe
86 connecting to the manifold 75 and a pipe 87 connecting to a
waste tank 88. The exposed portions of the valves must be inert and
are preferably stainless steel with Teflon inserts though other
materials may be employed if suitably inert to the solutions.
Manifold 75 is also connected through a pipe 100 with a pump
metering chamber generally indicated at 101. All parts of the
metering pump exposed to the solution should be relatively inert.
Teflon plastic and stainless steel have been employed for this
purpose. Chamber 101 is formed in a piston operated metering pump
which is designated 99 and in this case includes a reversible
electric motor 105, FIG. 7, connected to drive an internally
threaded tube member 106 which in turn receives a threaded piston
rod 107 such that when the armature of motor 105 rotates in one
direction piston rod 107 is moved forwardly to move piston 110
toward the cylinder port 111 and when the motor armature turns in
the opposite direction piston 110 is moved away from port 111.
Piston rod 107 mounts a pin 112 which moves back and forth in a
slot 113 located in bracket 114. Four micro-switches 115, 116, 117
and 118 are mounted in the path of retracting movement of pin 112
so as to strike pin 112 and be actuated. Switches 115, 116, 117 and
118 are arranged in the circuitry so that one of the switches can
be selected to control the amount of movement of pin 112 during
retraction by deenergizing motor 105 and by this means very
precisely control the amount of solution metered by pump 99 and
indicated by levels A, B, C and D. It should be understood that
appropriate circuitry "commands" motor 105 to turn to advance
piston 110 when a solution is to be metered, other circuitry
commands piston 110 to retract, other circuitry selectively
controlled by micro-switches 116-118 command motor 105 to stop and
other circuitry commands motor 105 to turn so as to advance piston
110 to pump out the metered solution. Annular piston edges 109 are
formed to assist sealing.
The processing chamber 90 of the present invention is generally
like but differs in an important respect from that shown in prior
U.S. Pat. Nos. 3,526,203 and 3,576,176. Specifically, the
solutions, instead of being admitted at the top through plural
inlets, are both admitted and discharged through a single port 91
in the bottom of chamber 90. Level sensing is thus avoided by means
of the described metering pump. Chamber 90 has a tapered bottom
which assists in dumping solutions and also allows a relatively
small number of capsules to be processed. To describe the
processing chamber 90 in more detail it will be seen that the
removable cap 120 is provided with a clear glass window 121.
Chamber 90 is physically located within cabinet 29 near
refrigerated compartment 40 which minimizes solution precipitation,
particularly that of osmium tetroxide. Cap 120 is positioned
slightly above the horizontal cabinet wall 123 which provides a
relatively large unobstructed work surface to the side of
processing chamber 90. The chamber should be inert to all solutions
and has been made of Teflon plastic. Stainless steel and other
materials may be used. As shown in FIG. 8, a small vent hole 124 is
provided on the upper inner wall of the processing chamber which
allows the solutions to drain and also allows the fumes from
solutions to be exhausted through an outside hole 125 by an exhaust
fam 130 connected through a flexible conduit 131 to a suitable
hood, not shown. That is, any fumes escaping through holes 124, 125
are drawn into the interior of cabinet 29 and then exhausted by fan
130. Also, when cap 120 is removed any fumes escaping from chamber
90 are drawn into apertures 126 in top wall 123 as shown in dotted
lines in FIG. 1. The fumes are then drawn inside cabinet 29 and
exhausted by fan 130. During processing the capsules are secured in
a tissue capsule rack of the type described in U.S. Pat. No.
3,576,176, indicated at 92, FIG. 8.
The processing of tissue particles according to the invention has
been described as generally involving placing the tissue particles
in the receptacles; placing the receptacles in the receptacle rack,
placing the rack in the chamber and then drawing the solutions into
the processing chamber in some predetermined time sequence. In this
regard the present invention elminates level sensing in chamber 90
and allows the amount of each fluid drawn into the processing
chamber to be precisely metered according to the number of
receptacles being processed. Thus, the panel board in FIGS. 5 and
10 will be seen to have a volume control switch 132 marked A, B, C
and D which correspond in one embodiment to respective loads of one
receptacle, (10ml.), two to four receptacles (25 ml.), five to 28
receptacles (50ml.) and 29 to 48 receptacles (100 ml.). Switch
positions A, B, C and D effectively "select" a corresponding
micro-switch from the group of pump microswitches 115, 116, 117 and
118 and thereby control the level of fluid used of a particular
solution. Additional program circuitry, not shown, "commands" the
pump to otherwise start and stop as required.
The circuitry also includes what is called a "cycle control" which
is shown at 135 in FIGS. 5 and 10 and which allows either 1, 2 or 3
solution changes of each stock solution. That is, the group of ten,
three position switches shown in FIG. 5 and 10 at 135 are suitably
connected into the illustrated program control circuitry so that
each solution can be controlled during a particular program as to
the number of changes 1, 2 or 3 of that solution. Thus, the cycle
switch for solution 1 may be set to call for solution 1 to have one
change, the cycle switch for solution 2 may be set to call for
solution 2 to have two changes, the cycle switch for solution 3 may
be set to call for solution 3 to have one change and so forth, as
shown in FIG. 10. Furthermore, cycle switches may be set one way
for one program and immediately after that program is terminated
the switches may be moved to a different array for the next
program. In this manner, a versatility not heretofore achieved is
provided. Those skilled in the art will, of course, understand that
the cycle switches by their settings control whatever program
circuitry is employed. FIG. 10 is intended to simply illustrate one
such configuration based on using a stepping switch and standard
logic type circuits.
In another aspect of the programming, the present inention also
provides for a so-called "time control." The time control operates
on the basis that each program is made up of some multiple of a
standard base time, e.g., 15 minutes. A switch 150, shown in FIGS.
5 and 10, is a multiple position timing switch and each position
corresponds to a portion of the base time period of 15 minutes.
That is, switch 150 can be set for any time from zero to 15
minutes. In addition to switch 150 the time control also includes
for each of the 10 solutions a multi-position "time" switch
indicated at 155, which can be moved to a position to indicate
either one, two, three, four, five, six etc., base units of time.
Thus, if the timing switch 150 is set on ten minutes and the
particular time switch 155 for solution 1 is set on the 2 position
this would mean that solution 1 would stay in processing chamber 90
for 20 minutes (two times the base 10 unit) and then run out. That
is, each cycle with solution 1 would be for 20 minutes. If the
corresponding cycle control switch 135 for solution 1 were set on
its 1 position the exchange of the solution 1 with the particles
would only take place once but if, for example, the cycle control
switch 135 were set on, say, its 2 position the exchange would be
repeated for another twenty minute period. The base time control
circuitry as well as the cycle control circuitry is, of course,
connected to suitable programming circuitry as generally
illustrated in FIG. 10. Thus, any solution can be regulated
independently of all other solutions as to the number of cycles and
the time per cycle merely by positioning the switches 135, 150 and
155.
The circuitry of the invention also incorporates a set of ten,
three position, hold-pass switches 170 having positions indicated
by "N" for normal, "P" for pass and "S" for stop. Switches 170 are
suitably connected into the program circuitry as indicated in FIG.
10. As background information it may be noted that tissue
processing of electron microscopy size tissue is subject at times
to many collateral operations for experimental purposes or to get
certain results supplementary to those achieved by routine
processing. That is, in some cases a program can be set up which
simply calls for the tissue particles to be processed step by step
through all the solutions. In other cases however the research
investigator may, for example, want to bypass certain solutions or
he may want to, at least temporarily, terminate the program after a
particular solution. When the particular hold-pass switch 170 is
set on "N" for normal the solution is used in a normal way and the
switch position has no effect on the program. However, when a
selected hold-pass switch 170 is set on "P" this switch setting
directs the program circuitry to pass the respective solution in
the program and not use it. Thus, if the hold-pass switch 170 is
set on "P" for solution 3, this solution would not be used at all.
Also, if the particular hold-pass switch 170 is set on "S" the
associated program circuitry is so arranged as to call for the
program to shut down immediately after such solution has been
dumped. Thus, if the operator technician is directed to stain all
the particles in a special stain solution between, say, solutions 6
and 7 the technician may set the hold-pass switch 170 corresponding
to solution 6 to the "S" position and the program will temporarily
stop after solution 6 has been dumped to allow this collateral
staining operation.
Considering further the circuitry and control panel shown in FIGS.
5 and 10, there is also incorporated a selector switch 160 having
ten positions, corresponding to the ten solutions, and which allows
the operator of the processor to select as a starting point in any
program any of the ten respective solutions. Thus, a particular
tissue may require special processing, in only, say, solutions 7,
8, 9 and 10 and this can be immediately obtained dependent only on
a proper initial setting switch 160.
To complete the general explanation of the circuit it will be
understood that given the concept of providing switch means to vary
the base time, the cycles per solution, the particular solution
starting point, the time per cycle per solution and processing
chamber level that many and varied logic, stepping switches and
other program and timing arrangements other than that shown
generally and substantially in FIG. 10 may be employed in
conjunction with such switch means. That is, the 10 cycle switches
135, the 10 hold-pass switches 170, the 10 time switches 155, the
selector switch 160, the volume switch 132 and the base time switch
150 may be used to control a variety of program circuits suitable
to the invention and well known in the art of programming. The
circuitry also includes a start switch 180 whose purpose, and
circuit arrangement, is to start a selected program once the
various program selected switches have been positioned as required
for such program. In addition there is provided a release switch
181 and a so-called "clean" switch 182. The purpose of release
switch 181 is to provide a means to operate the dump valve 85 so
that in the event of a mulfunction or for other reasons it becomes
necessary to immediately drain chamber 90 this can be done by
actuating release switch 181. Switch 181 is wired accordingly to
override all other program commands. The clean switch 182 serves
another useful purpose and is wired in the circuitry so as to
command only certain solution control valves 73 to operate in some
predetermined sequence designed to flush or clean out the chamber
90, manifold 75 and the various passages at the end of one program
and before another program starts. Thus, clean switch 182
effectively commands a subsidiary program intended for cleaning the
system and not for processing as such. In one embodiment clean
switch 182 and the associated circuitry are programmed to dump
fluid from chamber 90, then to rinse twice with propylene oxide,
solution 9, then once with absolute alcohol, solution 8, then to
dump and dry. Indicator lamps 95, 96 and 97 indicate as shown in
FIG. 10. Lamp 94 indicates a waste tank full condition.
It may be mentioned that an alternate manifold arrangement has been
employed experimentally with level sensing as distinct from
positive metering. In the alternate arrangement the manifold
allowed the solutions to flow by gravity and entry was made through
a single port located at the top of the processing chamber,
however, this was found not to offer the advantages of the present
invention. The illustrated manifold of the invention has proven
particularly advantageous in providing a means for shortening flow
passages and for minimizing intermixing of solutions. It also
provides a means for mounting the solution control valves, the dump
valve and the processing chamber valve as well as the metering
pump. The use of a single solution entry and discharge port in the
processing chamber reduces possible points of contamination from 10
to 1. Thus, while the method and apparatus of our own prior art
worked and provided a highly significant advance in the art it can
be seen that the present manifold arrangement has further advanced
the art and in novel and significant respects.
Agitation and vibration of the solutions during processing to
increase solution penetration through the receptacle pores has been
tried but is generally not deemed necessary. Also, the required
agitation or vibration apparatus has not been found to be justified
by the results obtained. However, it is, of course, recognized that
vibration or agitation may be employed and may in the future, as
automatic tissue processing knowledge becomes more fully
developed,prove justifiable in certain tissue techniques. Thus, a
vibrator "V," FIG. 8, may be employed to vibrate chamber 90 or a
motor "M," FIG. 8, may be employed to rotate rack 92 during
processing somewhat like an electric ice cream freezer dasher to
obtain solution agitation.
While not shown, the control panel and circuitry may include,
depending on the specific program circuitry employed, a "set"
switch to set the program logic circuitry once the external control
switches have been selectively manually positioned and a "reset"
switch to restore such logic circuitry to a particular program
state. Such "set" and "reset" switches have been employed in one
experimental embodiment and a starting sequence used which used the
solution indicator lamps shown in FIG. 10 to indicate to the
operator that a selected program has been "set" and is ready to
start on a particular solution indicated by the associated solution
indicator lamp, in solution indicator lamp bank 140.
What is deemed most significant and most important to providing a
reliable day-to-day automatic electron microscopy processor is the
achievement in this invention of a practical system of measuring nd
minimizing fluid contamination. Also, of special importance to the
present invention has been the achievement of a manual switch
selector system that readily adapts to any of many well known forms
of program circuitry to allow selection of base time, number of
solution cycles, starting solutions, positive level measuring,
particular solutions for particular processing and cleaning
programs, and alternate terminal solutions for particular programs.
The research investigator and histology technician is thus provided
with a new apparatus and method for both duplicating hand
processing procedures and, more importantly, for allowing
completely new processing techniques not heretofore known in the
art.
While primarily directed to electron microscopy size tissue, it is
recognized that much of the fluid exchange and timing apparatus of
the invention lends itself to automatic slide tissue staining as
welll as to light microscopy processing directed to processing 3 to
5 millimeter thick pieces of tissue. For staining slides the
processing chamber of the invention may be in the form of a
rectangular well and used in conjunction with a conventional
staining slide tray adapted to hold, for example, 60 3 .times. 2
slides. The staining fluids would be recirculated rather than
dumped but would be periodically changed depending on the amount of
use. The solenoid valves and pump operation could be controlled
accordingly and such staining could be performed at room
temperature. For light microscopy work larger solution containers
should be employed and all lines and pump surfaces handling
paraffin should be maintained at a temperature high enough to keep
the paraffin in a fluid state. The tissue receptacles for such
ligth microscopy processing could be of the type shown in U.S. Pat.
No. 3,411,185 and a plurality of such receptacles could be stacked
in a perforated basket for moving into and out of the processing
chamber. The processing chamber could be the same rectangular well
form used for staining thus adding to the versatility. As with
staining the fluids would normally be recirculated and selected
fluids periodically changed depending on the amount of use.
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