DE2416661A1 - DEVICE AND METHOD OF EMBEDDING A SPECIMEN - Google Patents

Description

t 2 4 I O D D J

Dr. - Ing.H / KS RUSCHKiI

S 1538 Dip! .- i, fci. CL-F RUSCHKE Munich, April 1974

Dipllng H ^Λ NS E RUSCHKE

Dipl.-Ing. H · ^Λ NS E. RUSCHKE

8 MUNICH ϊίΟ
Pienzenauersfaße 2

SHERWOOD MEDICAL INDUSTRIES INC., St. Louis, Missouri 63103

Apparatus and method for embedding a test piece

The present invention relates to devices for embedding specimens and methods of forming media blocks in which embedded specimens are contained and from which specimen portions can be cut for analysis.

In order to be able to place a test piece, e.g. a histological tissue sample, on an objective carrier for microscopy, the sample is generally embedded in a block of an embedding medium, e.g. in paraffin, and then individual parts or slices of the block and the test piece are passed through the block a cutting device, for example by a microtome, cut from the block. Previously, the specimen was placed in the bottom of an open-topped mold and molten paraffin was poured into the mold to cover the specimen. After the paraffin had solidified into a block, it became the

509807/0990

Taken out the mold and sliced through a microtome to get specimens that are on Lens carriers for microscopic examination can be placed. In U.S. Patent 2,996,762 a method is described, according to which a lower molding from the solidified paraffin, which the specimen is removed while a top molding with the paraffin remains in place and for clamping is used in a microtome. The method according to this patent specification, however, requires the body to be handled embedding medium, such as paraffin. This, in turn, requires melting paraffin and standing by of equipment parts which are relatively expensive, it requires pouring and dispensing of molten paraffin the material is placed in every single mold set up in a hospital or laboratory that contains a specimen the influence of a foreign matter and generally requires careful and experienced technical Skilled workers. Such processes also generally require excessive heating of the paraffin and besides a laboratory cleaning.

It is therefore an object of the present invention to provide an apparatus for embedding specimens and a To provide a method for embedding test pieces in which the aforementioned disadvantages are substantially avoided will.

509807/0990

Another object is to provide an improved apparatus for embedding the samples that does not require casting still requires dispensing of liquid paraffin at the point of use.

Another object is to provide a reservoir for the sample embedding material and a molding apparatus to create that is easy to use and not auxiliary equipment for melting, pouring and dispensing required to obtain a test piece embedded in a mass of potting material.

Another object of the present invention is to; Specimens a new embedding process in embedding media blocks for cutting slices to create Obtain test pieces for investigation purposes, which generally meet the requirements for skill and reduces the time required and eliminates the cleaning difficulties associated with previous methods usually occurred.

Another aim is to create a pre-filled supply pack with a predetermined amount of hardened, specimen-embedding material, with at least parts of the supply amount being used as melting parts in order to produce a molten mass of material with a material embedded therein.

- 4 -509807/0990

to deliver embedded specimens.

According to an embodiment of the present invention, a storage container for embedding material of Specimens and a molding device have been created which includes a mold which is formed from an upper mold part with an open upper end and a lower molding which has a closed lower end. In the The mold contains a solidified mass of specimen-embedding material which is meltable within the mold is when heat is applied to it, and the lower mold part is after the embedding material has solidified removable and provides a protruding lower portion of the solidified embedding material which the specimen contains. According to another embodiment, the upper mold has a removable one at its end Cover covered to close the interior of the mold and the solidified mass of the embedding material against foreign influence. In accordance with another embodiment of the invention is a method of embedding a specimen provided in which a mold is provided with upper and lower parts in which one has solidified Bulk of fusible specimen potting material is contained, as is heat input to melt the therein embedding material contained, introducing a test piece into the molten material and its Moving further into the lower molding, the solidification of the

- 5 509807/0990

Material within the mold and removing the lower mold part to form a solidified lower part of the embedding material with the sample embedded therein to expose.

These and other goals and benefits of the present. Invention are apparent from the following description and drawing.

Show it

1 is a perspective view of a storage container for sample embedding material and a molding apparatus according to an embodiment of the present invention

Invention,

FIG. 2 is an exploded view of FIG. 1 Arrangement,.

Fig. 3 is a cross section to the left of the line 3-3 of

Fig. 1,

FIG. 4 is a view similar to FIG. 3, but in the representation a process step of embedding a test piece in the embedding end

Material,

- 6 509807/0990

Fig. 5 is a plan view showing a block of the

embedding material with a test piece embedded therein and in a Shows microtome inserted, and

6 is a perspective view of a disk of the block of embedding material; which is shown in FIG.

In Fig. 1 is a storage container for sample embedding material and a molding device 10 with a mold 12, which has a peelable or removable lid 14 attached to the end of the mold. The mold 12 includes a upper mold part 16 and a lower mold part 18. As shown in FIG Figures 2 and 3 can be seen better, the mold 12 is previously filled with a predetermined amount of a solidified, the sample of embedding material 20. The embedding material 20 is usually made of paraffin, as is conventionally used for embedding test pieces, e.g. in a process, parts of tissue samples for microscopic examinations, e.g. for histological examinations.

The upper mold portion 16 is shown in a generally rectangular shape with two pairs of opposed parallel side walls 22 which define a generally square channel which is at both the top

- 7 -509807/0990

End and at the lower end is open (Fig. 3). The upper mold part is radially outward with a circumference protruding lower flange 24 is provided. The molding is relatively stiff and retains its in use Shape. It can be made of any suitable material, e.g. made of plastic such as polytherphthalate, polypropylene, or a relatively hard styrene, and may be made in any suitable manner depending on the material used, e.g., by injection or be shaped by bubbles. "~ ——" '

The lower mold part 18 is shown generally cup-shaped having four side walls 26 arranged in a generally rectangular shape and one im substantially flat bottom wall 28 which closes the lower end of the molding. The top of the molding is open and is provided on the circumference with a radially outwardly projecting flange 30, the lower mold part is shown connected to the upper molding 16, for example by means of a suitable adhesive 31 between the flanges 24 and 30. Preferably, the adhesive 31 is such that there is an adequate seal between the Molded parts 16 and 28 produced to prevent the embedding material or paraffin 20 in the melted State is leaking out of the mold, but it is not supposed to be the relatively easy separation of the upper from the lower mold part prevent. The adhesive 31 may be rubber-like, for example

. - 8 509807/0990

Adhesive or a rubber-based binder, which is preferably of a non-curing type. That Adhesive should preferably be able to hold the mold parts 16 and 18 sealingly together when the mold is exposed to the temperatures normally encountered during use of the device. It is desirable that the adhesive 31 has a low peelability on at least one of the molded parts 16 and 18 to to allow its removal. In some use cases the molded parts can also be set up so that they snap together or snap into place instead of an adhesive connection in a liquid-impermeable manner with or without a binder.

The lower mold part 18 is also provided with a handle or projection 32 which is connected to the flange 30 is and can consist of one piece with this, as shown in the drawing. The projection 32 is used as a means to grasp the lower mold part and quickly remove the molten paraffin 20 and the upper mold part 16 as explained below. The lower mold part 18 consists of a material whose shape at the usual temperatures during use occur, but which is sufficiently resilient to allow its relatively easy removal by pulling the projection 32 in a direction away from the upper mold part when the paraffin 20

509807/0990

is in the solid state. It can be made of any suitable material, such as the same material as Part 16, or of another suitable plastic material, or of a metal, e.g. aluminum foil. The lower part 18 is preferably designed so that it is more resilient or flexible than the upper mold part 16 ^ for the purpose of lighter Removal without harm to the melted paraffin, in the illustration it consists of a thin plastic material, which is significantly thinner than that of the upper molded part 16.

When assembling the reservoir for sample embedding material and the molding apparatus 10, the upper and lower mold parts 16 and 18 of the mold 12 are preferably connected to one another by the adhesive 31, and a solidified mass of embedding material 20 is inserted into the mold. For example, a predetermined amount of paraffin can be poured into the mold by pouring molten paraffin be introduced up to a predetermined height, namely up to a height indicated in the drawings is whereupon the paraffin cools and hardens. Optionally, a desired amount of solid paraffin can also be used placed in the mold.

The cover 14 can consist of a thin film, ___ ^^ - 'for example a flexible, elastomeric film, which is not

- 10 -

509807/0990

7416661

changed in an undesirable manner when the paraffin is heated to the melting temperature. The cover 14 can for example be a mylar or a vinyl film. The cover is attached by means of a suitable adhesive 33, which is made of the same material as the adhesive 31 can exist. The adhesive 33 can be applied to the upper end of the upper molding, as shown in FIGS Figures 1 and 3 can be seen. The cover 14 is shown with one made from the same piece Protrusion or handle 34 for its removal or detachment from the uppermost part of the upper mold part 16 in the To facilitate timing of use. The adhesive should allow relatively easy removal of the cover 14 from the upper mold part 16 without spilling the molten paraffin. The cover seals the mold so that the paraffin is stored in the sealed interior or chamber of the mold 12 and prevents foreign particles from entering the mold during storage, thus preventing that such particles penetrate the paraffin when it is melted. It is understandable manner of Importance of preventing any foreign substance from entering the mold, as this is what makes the microscopic examinations can disturb.

When embedding a test piece, e.g.

- 11 509807/0990

A piece of tissue for histological examinations becomes a storage container for the embedding material and the molding device 10 heated in a suitable manner to melt the paraffin. For example, the device 10 may be placed in a heated Atmosphere, e.g. in an oven, around the normally solid mass of the embedding paraffin to melt within the mold 12. The paraffin supply 20 in the fully filled mold 12 is so large that the paraffin in the molten state completely fills the lower mold part 18 and rises to a level in the upper mold part 16, in which the upper mold part carries the paraffin 20 when it later becomes a solid block or mass has hardened and the lower molding has been removed. E.g. the melted paraffin can halfway into the upper Rise molded part 16, as shown in FIGS is. The cover 14 is removed either before or after the wax has melted to the top of the part 16 and the mold 12 to open to access the interior of the mold and the paraffin therein to enable. That. Sample 38 in Fig. 4 is then placed in the molten paraffin and, as desired, aligned in the lower molding, and the paraffin can then cool and harden.

To avoid the risk of molten paraffin spillage, the cover 14 is removed or partially removed from the mold 12 only after the paraffin 20 has been removed

509807/0990 " ¹² "

melted within the container in which the mold is handled or agitated with molten paraffin will. The mold 12 is preferably placed on a heated surface or table, as at 39 in Fig. 4 shows the paraffin in the molten state during the above-mentioned alignment of the specimen to keep. After the desired alignment of the test piece, the cooling of the paraffin can be supported be that the mold 12 is placed on a cold plate.

After the paraffin 20 with the sample 38 embedded therein has hardened, the projection 32 and the Upper mold part 16 covered by hand and peeled off from each other to the lower mold part 18 of the hardened Remove the paraffin block and the upper molding. Since the paraffin reaches up into the upper mold part 16, it adheres to the side walls of the upper molding

so that the upper mold part the paraffin block 20 they! - ~ - '

holds on. Although the lower mold part 18 has sufficiently rigid side walls 26 to accommodate the paraffin and to shape, it is fairly flexible, as already mentioned above was to quickly from the lower end of the upper mold part 16 and from the hardened paraffin block without danger for the lower end of the paraffin block to be removed.

- 13 50 9 8 07/0990

When the lower mold part 18 is removed, that serves relatively rigid upper mold part 16 as a holder for carrying the solid paraffin block with the one embedded therein Specimen. The molded part 16 and the paraffin block 20 are placed in a microtome, as in FIG. 5 shown at 40, which may be in any suitable or known conventional form. The microtome 40 is shown with a pair of opposing chucks 42 and 44 attached to a pair of each other opposite, parallel, flat side walls, 22 of the right-angled molded part 16 rest in order to clamp the molded part firmly in the microtome. The shape will then placed exactly under a knife, shown at 48, which is passed through the molded paraffin block and the inside Cuts through the test piece.

In FIG. 6 there are shown a plurality of slices, indicated at 50, which have been obtained by the cutting operation indicated in FIG. Each disc contains a paraffin portion indicated at 20 ¹ and a specimen portion indicated at 58 '. One or more slices 50 can be cut from the paraffin block containing the embedded pattern. The block with the remaining paraffin and specimen mass can be saved for reference purposes if desired. Successive slices can be separated properly without breaking by using

- 14 509807/0990

flats 52 are provided at each corner of the disc; these flattened areas have arisen in that the lower molded part 18 is provided with flat corners 54 (FIG. 2) lying in the axial direction. The flattened areas 52 lying next to one another form incisions between parts 2O ¹ lying next to one another, which make it easier to separate the parts.

Each disc 50 can in a known manner on a conventional, transparent microscope lens mount (not shown) and, if desired, various treatments be subjected. The specimen on the disc can then be covered with a cover glass and placed in a Microscope can be inserted for microscopic examination.

The embedding device 10 thus serves as a pre-filled one Storage container or package for embedding material that also serves as a mold and provides an effective device for the method of embedding specimens. It is very important that the operating time be reduced and the device for treating and pouring the paraffin mass during embedding of the specimen is not needed due to the use of the present prefilled package and embedding device, and the embedding method described above is simple and effective. It is very important that the required Time for the embedding process described above

- 15 509807/0990

will often be greatly reduced compared to methods that involve handling large embeds of Epn for the specimen where the length of time can be critical when it comes to histological examination. at The present invention also, as noted above, avoids waste and eliminates the amount of cleanup that would normally be required required when a large amount of "paraffin" is used at the embedding station is substantially reduced. Furthermore, there is a risk that foreign particles or substances contained in the air in the embedding material penetrate, reduced by the fact that the embedding material supplied in the illustrated embodiment is sealed in the container chamber until the time of use by adhesive 33 and cover 14. In addition, is when using the device and the method according to the present invention, no measuring is required and there is also no inattention, since the supply pack 10 contains a desired, predetermined amount of material is.

- 16 509807/0990

Claims (9)

PATENT CLAIMS 1.j Storage container for specimen embedding material and molding device for use in preparing embedded specimens, with an upper storage container part which defines an upper mold part which is open at its upper and lower end and has axially lying side walls, with a lower storage container -Part that defines a lower mold part with a bottom wall, which has its lower end off-SCHUBt ₇ and an open, upper end, which is connected to the lower end of the upper mold part, with a mass in a predetermined amount of a solidified, meltable sample Embedding material within the reservoir and the molding device, and with a removable cover that closes the upper open end of the upper mold part, while the lower mold part is removable from the upper mold part and the embedding material when the embedding material is in the solidified state and a protruding lower end part of the solidified incl forms ettungsmaterials, which extends in the axial direction under the lower end of the upper molded part. 2. Storage container and molding device according to claim 1, characterized in that the upper mold part behaves- - 17 509807/0990 is moderately stiff compared to the lower molding. 3. Storage pack and molding device according to claim 1, characterized by a first adhesive layer which makes the upper and lower mold part liquid-impermeable and a second layer of adhesive that seals the cover against the top end portion of the top molding seals. . 4. Storage container and molding device according to claim 1, characterized in that the lower mold part on his the upper end has a flange which is attached to the lower end of the upper molded part, and a projection which extends laterally from the flange to facilitate removal of the lower mold part from the upper mold part. 5. Storage container and molding device according to claim 1, characterized in that the lower molded part has flat corners extending in the axial direction, whereby, after the solidified embedding material with the test piece contained therein is cut into slices, closely spaced panes have notches that make it easier to separate the panes without damaging them, and by an adhesive that sealingly connects the upper mold part to the lower mold part and a separation of the two molded parts from each other by hand. - 18 509807/0990 6. A method for embedding a test piece in an embedding material, characterized by an upper mold part with ends open at the top and bottom and a lower mold part with ends open at the top and closed at the bottom and a mass of solidified, fusible material containing a test piece, which is in the form is arranged by heating and melting the embedding material, then by inserting a test piece into the molten embedding material at the closed end of the lower mold part, by cooling and solidifying the embedding material, and by removing the lower mold part from the solidified embedding material, exposing a block attached to the upper mold part of embedding material, the lower part of which contains the test piece embedded therein at its lower end. 7. The method according to claim 6, characterized in that both the mold and the embedding material are heated at the same time. - -. · --- 8. The method according to claim 7, characterized in that that the heating of the mold with the embedding material contained therein takes place in a heated atmosphere. 9. Method of making part of a fabric sample for microscopic examination, characterized by the storage container for sample embedding material 509807/0990 " ¹⁹ " and molding apparatus according to claim 1, removing the cover, Heating of the storage container and molding device and melting of the embedding material contained therein, Inserting a test piece through the open end of the mold into the lower mold part while the Embedding material, aligning the specimen in relation to the plane of the bottom wall, then by cooling and solidifying the potting material within the mold. Forming a block of potting material with a test piece embedded in its bottom part, removal of the lower mold part from the solidified embedding material and from the top mold, clamping the top mold in a microtome and cutting off a disk from the bottom portion of the block of potting material in a plane generally parallel to the plane of the Bottom end of the block, the disc containing part of the test piece embedded in the block. 609807/0990 Blank