U.S. patent number 3,832,923 [Application Number 05/288,209] was granted by the patent office on 1974-09-03 for method for the production of microtome slices and device for the implementation of the method.
This patent grant is currently assigned to Schwarzer Company. Invention is credited to Gustav Lassmann, Peter Rieger.
United States Patent |
3,832,923 |
Lassmann , et al. |
September 3, 1974 |
METHOD FOR THE PRODUCTION OF MICROTOME SLICES AND DEVICE FOR THE
IMPLEMENTATION OF THE METHOD
Abstract
The invention pertains to a method for the production of
microtome slices, in which the slices, separated from the object,
reach a liquid bath, then are removed from the latter and are
prepared for further processing; further, a device for the
implementation of the method, with a microtome exhibiting a
stationary knife and an object slide (carrier) effecting the
cutting movement.
Inventors: |
Lassmann; Gustav (Vienna,
OE), Rieger; Peter (Walldorf, DT) |
Assignee: |
Schwarzer Company (Munich,
DT)
|
Family
ID: |
5832396 |
Appl.
No.: |
05/288,209 |
Filed: |
September 11, 1972 |
Foreign Application Priority Data
Current U.S.
Class: |
83/16; 83/28;
83/167; 83/170; 83/171; 83/915.5; 83/24; 83/152 |
Current CPC
Class: |
G01N
1/06 (20130101); Y10T 83/0472 (20150401); Y10T
83/283 (20150401); Y10T 83/0453 (20150401); Y10T
83/2185 (20150401); Y10T 83/222 (20150401); Y10T
83/0414 (20150401); Y10T 83/293 (20150401) |
Current International
Class: |
G01N
1/04 (20060101); G01N 1/06 (20060101); G01n
001/06 () |
Field of
Search: |
;83/15,16,24,152,170,171,915.5,167,28 |
References Cited
[Referenced By]
U.S. Patent Documents
Primary Examiner: Yost; Frank T.
Attorney, Agent or Firm: Wall; Joel Nealon; William C.
Berkenstack, Jr.; Howard R.
Claims
We claim:
1. A method for the production of microtome slices, in which the
separated slices from the object specimen reach a liquid bath, and
are then removed from the bath and prepared for further processing,
said method comprising the steps of:
a. preliminarily cutting the object specimen at a first cutting
station before the object specimen slicing step;
b. cutting the object specimen to produce object slices in
consecutive sequence and predetermined number so that the slices
adhere to each other on their edges forming a sectional strip;
c. maintaining a predetermined temperature at the blade which
performs the cutting;
d. floating the sectional strip on the liquid bath;
e. floatingly transporting the sectional strip to a carrier
immersed in the bath;
f. adhering the sectional strips to the carrier and removing
together therewith when a predetermined number of slices is
attained; and
g. thereafter conducting the object slices to further
processing.
2. The method according to claim 1 and wherein the method comprises
the further step of forming the objects in small blocks and storing
said blocks in a desired position, then conveying said blocks to an
object carrier, then placing said blocks in an object holder, and
then slicing said blocks.
3. A device for slicing object specimens comprising in combination
a microtome having a stationary knife, wherein a section of the
knife forms a first cutting station for making an initial pre-cut
prior to slicing the object specimen, means for positioning said
section for said initial pre-cut, an object holder for moving said
object specimen thereby effecting a slicing operation, a liquid
container mounted to the knife containing a bath on which the
object specimen slices are floated, means for controlling the
temperature of the knife and the temperature of the bath, means for
automatically removing the slices from the bath and for conveying
the slices to a carrier, and wherein said removing and conveying
means includes a lifting head for producing a suction force on the
slices.
Description
BACKGROUND OF THE INVENTION
For microscopic studies, thin sections (slices) of the objects are
required, which after corresponding subsequent treatment, such as
staining, can be microscopically studied. Whereas for the taking of
tissue specimens and for the facilitating of microscopic studies,
at least partially automatic methods were developed, heretofore
only manual methods have become known for the production of
microtome slices, aside from the fact that microtomes with motor
drive were developed for the cutting movement of the knife or
object slide.
By means of the invention, it is intended that the gap existing
between the known removal method and the known microscopy method be
bridged in such a manner, that a microtomy method is offered for
the production of sections, which is appropriate for automatic
operation. Further, it is intended that suitable equipment be
offered for carrying out this method.
Starting with the fact that in microtomy, it is likewise known how
to let the slices slide, after separation from the object by the
knife, into a liquid bath, or to cut directly in liquid, the task
presented in regard to technical method is so resolved, that the
slices are separated from the object in consecutive sequence, and
float on a liquid bath, then are removed from the latter, conveyed
to a carrier slide, and thereupon conducted to further processing,
as for instance a staining process.
This method can also be so further advantageously developed, that
in a consecutive sequence several slices each time, in a
predetermined number, are separated from the object, which adhere
to each other on their front surfaces and form a sectional strip;
and then the sectional strip, floating on the liquid bath, is
removed from the fluid, conveyed to a sectional strip
slide-carrier, and conducted to further processing. When the slices
floating on the liquid bath, or the sectional strips consisting of
a predetermined number of slices adhering to each other, are raised
up out of the bath and laid on the given slide-carriers, this can
appropriately take place in such a manner that the sections or
sectional strips, with the slide-carriers are removed together from
the bath, the application of the slices or sectional strips onto
the given slide-carriers taking place directly before, or during,
the removal from the liquid bath.
A device for the receiving of the slices already enjoys technical
status. This known device utilizes a moving belt as a
slice-receiving element, which with intensive directional shift, is
conducted upward onto the knife back, and in so doing, receives the
slices derived from the knife. In the present case, such a device
would be inappropriate, because it is not readily insertable in the
flow.
The method according to the invention can also be so characterized,
that the objects occurring in the form of small blocks or the like,
in which the actual specimens are embedded, are provided, before
the cutting, with two inclined surfaces or initial faces bordering
the cutting surface having edges running parallel to each other,
and are so held during the cutting by the object holder, that the
aforementioned edges run essentially parallel to the knife cut.
A further important feature of the invention consists in cutting
the object under defined temperature conditions in the knife blade.
The slices can also be advantageously extended, directly after the
cutting by means of fluid, or a temperature-controlled liquid bath
on which the slices float, regardless of whether individual
sections are involved, or sectional strips consisting of several
slices of predetermined length adhering to each other. Again, in
further development of the invention, the removal of the slices or
sectional strips out of the liquid bath, can take place in
conjunction with the achievement of a predetermined number of
slices or predetermined sectional strip length.
In comparison with the present state of the technique, the
essential difference in the method presented according to the
invention consists in the fact that all the steps of the method are
amenable to automation and can proceed in a positive sequence. It
has likewise proved advantageous in reference to the task set for
the invention, to the extent that the slide-carriers loaded with
slices or sectional strips of a predetermined number or length,
after drying of the slices if necessary, as well as objects
occurring in the form of small blocks, can be stored in greater
number in desired position. The small object-blocks can then be
individually removed, one after the other, from the supply,
conducted to an object slide-carrier, taken from the latter and
then cut, whereupon the storing of the small object-blocks takes
place, appropriately in a corresponding storage container.
The method given above thus creates all prerequisites for an
automatically-functioning operation in the production of microtome
slices, which begins with the storage, if necessary, of small
object-blocks previously prepared, and extends over the cutting,
extending and laying of the slices or of sectional strips formed of
individual connected slices, onto slide-carriers, for instance
small, thin glass laminas, up to the point of storing the
slide-carriers loaded with slices or sectional strips.
To resole the task further involved in the invention, it is
intended that a suitable device be described for the application of
the method which, based on microtome exhibiting a stationary knife
and an object slide-carrier effecting the cutting movement, is so
characterized that a liquid container with a liquid bath on which
the slices separated from the object float, is attached to the
knife, and that a device is planned for the automatic removal of
the slices from the liquid bath, and for the transfer of the slices
to the carrier-slides receiving the latter.
A device of the type indicated here can also advantageously exhibit
an initial coordinated cutting or pre-cutting station, adjoining
the actual cutting station, with the section of the knife located
in the liquid bath attached with the knife, so that when initially
cutting a small object-block, the resultant pre-cut slices do not
go into the liquid bath, and into the further processing procedure.
The implementation of such an initial cutting station can be
characterized by a knife holder carrying the knife and a liquid
bath extending over a portion of the knife width. The knife holder
is constituted as a carriage which can be shifted along the
knife-blade edge facilitating a knife displacement in such a manner
that the knife section coordinated with the initial cutting station
is located in the path of movement of the object, while after the
pre-cutting, the knife is displaced by means of its displaceable
holder, parallel to the knife blade edge, until the knife section
is coordinated with the liquid bath and the main cutting
station.
Naturally, within the scope of the present invention, the knife can
be stationarily arranged with two adjoining sections, one each of
which is coordinated with a pre-cutting station and a main cutting
station. In which case, however, it is necessary to have a movable
object slide-carrier so that the object may be movable between the
knife sections coordinated with the pre-cutting station and the
main cutting station.
The device according to the invention can also be so characterized,
that the device can be coordinated with the liquid bath and at
least the knife section coordinated with the main cutting station,
for the cooling or heating of the knife and for the temperature
control of the liquid bath. By so doing, required conditions are
fulfilled for many specimens under defined temperatures at the
knife blade, and a defined temperature control of the liquid bath,
whereby a waving or crimping of the slices which is practically
unavoidable during the cutting, is removed. The slices are
therefore extended and thereby placed in a form which is
serviceable for further processing.
Finally, the device indicated here can also be further
characterized by an apparatus for the removal of the slices from
the liquid bath, and for the placement of same on carrier plates,
which exhibit a lifting head which is movable between the surface
of the liquid bath and a slice delivery station, which head is
closed on its underside by a perforated supporting plate capable of
admitting an indraft (vacuum). Differing from the previously
mentioned form of exemplification, the apparatus for the receiving
of the slices from the liquid bath can also consist of a mounting
receiving a carrier plate in each case, which mounting is so
controlled that the carrier plates are immersed in the liquid bath,
are brought into contact with the slices floating in the bath, and
together with the slices are removed from the bath, whereupon the
storing of the carrier plates takes place. The carrier plates can
be brought into contact with the slices in such a manner that by
means of the mounting, they are immersed in the liquid bath and
removed, in the course of which, at about maximum immersion depth,
the end of a slice or sectional strip located at a distance from
the knife comes into contact with the given carrier plate, and the
raising onto the carrier plate takes place when removing same from
the bath through adhesion. Anothe possibility consists in bringing
the carrier plates by means of the mounting, under the slices
floating on the liquid bath, and to receive these slices, during
the raising movement of the carrier plates, onto the latter. The
device can then further possess a plate supply container, receiving
the carrier plates from the mounting, in which container the
introduction of the carrier plates takes place by means of the
mounting for the carrier plates.
For the implementation of an essentially automatic operation of the
method according to the invention, for the production of object
slices, the device can also possess a block supply container, which
receives prepared small object-blocks in a predetermined number for
cutting, with which the object slide-carrier is loaded one after
the other. Further, a block container can be provided, to receive
the small blocks after the cutting.
Below, it is intended to clarify the invention in greater detail,
reference being made to the attached drawings, which on the one
hand show the flow of the method and on the other hand, the
essential features of the device without, however, establishing any
restriction. The following are show in schematic views:
FIG. 1, the flow of the method, reference being made to a block
diagram;
FIG. 2, the basic construction of the device, in agreement with the
diagram according to FIG. 1;
FIG. 3, a cutting station with a tank receiving the liquid bath,
and the knife;
FIG. 4, a cross-section through knife and tank, along the line
IV--IV in FIG. 3;
FIG. 5, a cross-section through knife and tank showing the removal
of the slices or sectional strips out of a liquid bath, with
simultaneous conveyance onto a carrier plate;
FIG. 6, a cross-section through knife and tank showing removal of
the slices or sectional strips out of the liquid bath;
FIG. 7, functional schematic showing the removal of the slices out
of the liquid bath by means of indraft (vacuum);
FIG. 8, functional schematic showing the placement of the slices
onto a carrier plate; and
FIG. 9, schematic representation of the preparation of the objects
occurring as small blocks.
According to FIG. 1 and FIG. 2 of the invention, the small
object-blocks 10 prepared for cutting are received by a block
holder 11, and from the latter, one after another, reach a
microtome 13 in the direction of the arrow 12. There, the small
blocks are received by the holder 14 of an object slide-carrier 15,
which executes a down-and-up directed cutting and return movement
(double arrow 16 in FIG. 2) as well as the infeed movement, not
further represented here, to a stationary microtome knife 17 and a
corresponding return movement on the return stroke. After the
cutting, the sliced objects from the object slide-carrier 15,
according to arrow 18, reach a block storing container 19, and are
kept in the latter. In each case after the release of a sliced
object block, and its placement in a block supply container, an
unsliced object is conducted to the object slide-carrier, and is
taken up by its holder. The control of the device takes place by
means of a central control element 20.
Parallel to the object conveyance to the object slide-carrier 15,
and for disposal of the small object-blocks sliced, there takes
place, likewise controlled through the central control element 20,
the conduction from a carrier holder 24 of lamina carriers 25 in
the direction of the arrow 26 to the slicing station; the placement
of the slices onto the carriers at 27 and, with insertion of a
drying process at 28, the disposal of lamina carriers loaded with
slices, in a supply container 29 (indicated by arrow 30), so as
then to be conducted to further processing, for instance a staining
procedure, or else subjected to the stored position. The
transistion of the slices from the slicing station onto the lamina
carriers 25 is identified in FIG. 1 by arrow 32.
As FIGS. 2 and 3 especially show, the knife 17 of the microtome 13
is taken up by a knife holder 35, which in turn is arranged on a
guide plate 36, and in longitudinal direction of the knife is
displaceable according to the double arrow 37, clear across to the
cutting movement of the object slide-carrier 15 and paralle to the
knife blade edge 38 without modification of the slicing thickness
which was set. About half way on the width extension of the knife
17, a stationary liquid container 40 is attached to the knife back,
hence on the side most distant from the object slide-carrier 15,
arranged on the knife holder and hermetically sealed against the
knife back. The knife section 41 loated in the area of the liquid
holder is coordinated with the slicing station itself, while the
section 42 located nearby forms the precutting (first cut) station.
The knife can be so adjusted through displacement of the knife
holder 35 in the direction of the double arrow 37, that either the
knife section 42 coordinated with the precutting (first cut)
station, or the knife section 41 coordinated with the main slicing
station, are located in the path of movement of the object received
by the object slide-carrier 15.
According to the details which can be seen in FIG. 4, the liquid
container 40 is constructed from a tank, whose bottom extends,
joining the knife back, at only slight depth from the latter, and
then runs steeply downward. In the vicinity of the knife blade, the
liquid bath 44 received by the liquid container 40 therefore has
only slight depth. Between the knife back and the bottom coursing
steeply downward, attaching to the tank section of slight depth, a
hollow space is located, in which the cooling and heating element
45 is located, which it not important to specify here in detail. On
the adjoining side of the container wall, a further heating element
46 is arranged. The cooling and heating device 45 and the heating
device 46 serve, on the one hand, for the adjustment of the cutting
blade 38 to a defined temperature, and on the other hand, for the
heating up of the liquid bath 44, likewise at a defined
temperature. In a model developed, a cooling of the knife took
place, by means of the device 45, at about -10.degree. c, while the
liquid bath 44 was heated to about +50.degree. C. In so doing, a
stable cutting temperature of +10.degree. C was established on the
knife blade. The temperature-influencing of the knife blade by the
higher bath temperature was slight, due to the slight liquid depth
in the vicinity of the knife blade. A slice or a sectional strip
flowing from the knife back (identified by 48 in FIG. 4) thus
reaches, leaving the knife blade, the area of the liquid bath which
is gradually becoming warmer. The slice (or sectional strip)
floating on the liquid bath receives, in so doing, a surface
extension which is advantageous for further processing.
FIG. 5 shows, in simplified representation of the liquid container
40 with the liquid bath 44, a slice 48 floating on the latter,
which is removed from a lamina carrier 25, vertically immersed in
the liquid bath 44, by means of a mounting 50. The slice or the
sectional strip 48 attaches, with its edge which was distant from
the blade edge 38 of the knife 17, to the lamina carrier 25 due to
adhesion, so that during the removal taking place in the direction
of the arrow 51, the sectional strip lies flat on the lamina
carrier 25. The further movement of the lamina carrier 25 is so
controlled by means of the mounting 50, in a known manner which is
not further represented here, that the lamina carrier 25 is
inserted, in the manner demonstrated in FIG. 2, in a plate supply
container 29.
In FIG. 6, a monting 50' is demonstrated, which receives a lamina
carrier 25 in a position slightly inclined against the horizontal.
When in operation, the mounting 51 immerses the lamina carrier 25
into the liquid bath 44, whereupon, after reaching a predetermined
section strip length, the cutting process is interrupted for a
short while and the lamina carrier 25 is removed out of the liquid
bath in the direction of the arrow 51'. In so doing, the placement
is effected of the sectional strip 48, which was floating in the
liquid bath, on the end of the lamina carrier 25 most distant from
the mounting 50, and in this manner removed from the liquid bath.
After this, the placement of the lamina carrier 25 into a plate
supply container is effected, in the same way as with the device
described in conjucntion with FIG. 5. Before placement in the plate
supply container, the sectional strip or else a single slice, can
be subjected to a drying.
In FIGS. 7 and 8, finally, a further possibility is represented of
the slice or sectional strip removal from the liquid bath 44, as
well as of the slice or sectional strip placement onto a lamina
carrier 25. This device consists of a lift-head 55, which is closed
on its underside by a perforated holding plate 56 amenable to
indraft (vacuum) which is displaceable as demonstrated in FIG. 8
for the placement of the slices or sectional strips onto a lamina
carrier 25. The fact that during the placement of the slice or
sectional strip taken up by the suction head onto a lamina carrier,
the suctional effect is removed or the slice or sectional strip
must be conveyed through a short air-pressure blast, can be readily
understood by any technician.
FIG. 9, finally, demonstrates a possibility for the processing of
the small object-blocks in such a manner, that the latter after
being taken up in holder 14 of the object slide-carrier 15, exhibit
two edges, parallel to each other, running at right angles to the
cutting surface. The block processing can take place in a simple
manner by means of two knives 58, 59, which provide the small
object-blocks, arranged in series, with initial faces 60, 61,
running toward each other at about 90.degree..
As was already initially clarified in the description of the
figures, the small object-blocks 10 provided with initial faces 60,
61 go from the block supply container 11 into the object
slide-carrier 15, and are subjected to tension by the latter in a
manner which is known, but however is automatic. The knife 17 is so
adjusted (set), through corresponding displacement of the knife
holder 35, that for the formation of a pre-cutting (first cut)
station, the knife section 42 is located in the vertically flowing
path of movement of the object displaced in the direction of the
double arrow 16. In this position, the object block, placed under
tension, is pre-cut. After the completion of the initial cutting
process, the knife section 41, which is coordinated with the actual
cutting station, is shifted in the path of movement of the small
object-block taken up by the holder 14, and the slices occurring
during the cutting flow away from the knife blade into the liquid
bath 44, to float on the latter, as demonstrated by 48 in FIG. 4.
The slices or sectional strips are removed from the liquid bath, in
the manner described above in conjunction with FIGS. 5 to 8, and
stored, possibly after prior drying, on lamina carries 25, resting
in the plate storage container 29. This is clarified with arrows
30, 65 in FIG. 2. In the exemplification shown in FIG. 2, and
before the storing of the lamina carriers loaded with slices or
sectional strips in the storage container, a 90.degree. rotation of
the laminas takes place, along their longitudinal axes, as
indicated by the curved arrow 66. After the storing of the lamina
carriers, loaded with slices or sectional strips, in the storage
container, they are passed along in the direction of the arrow 67
in FIG. 2, to processing which is not represented here.
* * * * *