JPS58105061A - Preparation of sample of cellular tissue and device used for said method - Google Patents

Abstract

(57) [Summary] This bulletin contains application data before electronic filing, so abstract data is not recorded.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention provides freeze imprl, ntJ
The present invention relates to a method for preparing a cell tissue specimen by the method of n5) and an apparatus used therefor. Hereinafter, the apparatus of the present invention may be referred to as a freeze layering apparatus (oryoappooitor).

Conventionally, diagnosis of organ tissues, etc. has been performed using cryo-cryotomy (free cryosection).
zing aryotome). A section sample of tissue (tj-ssue) from a patient's diseased area (
5section) is removed by the surgeon, which is then first frozen by a pathology examiner (autopsy examiner), further cut into 71 [sections], and finally colored into microscopic specimens. Provided for diagnosis.

There is an alternative (but inaccurate) method to the one on paper. That is, the method involves simply depositing a section sample removed by the surgeon.
), but in most cases it is due to the mechanical stress caused by the sliding that the small cell material undergoes, resulting in the destruction of a portion of the tissue.
The conditions are inferior.

When the conventional method was carried out, the existence of DO82704300 was revealed. In the conventional method, a small cylindrical object of the autopsy specimen is cut with a knife in an appropriate apparatus α, and the surface of the small cylindrical object is cut. After cutting, the cylinder is immediately immersed in a freezing liquid (for example, liquefied nitrogen) to harden it.

The resulting sample has a considerable thickness and the surface under test is damaged due to the abrasion caused by the knife.

Apparatus for cooling autopsy specimens (see British Patent No. 102S97), apparatus for cooling autopsy specimens on microscope slides (see French Patent No. 1405270) or apparatus for cooling microscope slides (see United Kingdom Patent No. 102S97); Special #'l
However, it is clear that the purpose and function of the device of J'L et al. are completely different from those of the present invention.

By using the method and apparatus of the present invention, the drawbacks of the two methods mentioned above can be overcome. That is, by using the method and device of the present invention, histological analysis in a single layer becomes possible. The device of the present invention is highly reliable, accurate, and thin (thin) for microscopic regeneration of normal or diseased organs.
This is based on the basic concept that ``inner'' and ``('' refer to layers of cells observed under a microscope.In fact, when observing autopsy specimens, it is ideal to examine a single cell layer that accurately reproduces the structure. It is true.

In this method G, the substance (parθnchyma)
, that is, the cells of epithelial tissues and tissues that control special functions of other organs are shown as positive θp.
icturθ), and the stroma forming blood vessels, cell tissues, and some cranes can be observed as negative drawings (nogatj).
va piture). Discipline is objectively observed.

According to the present invention, the above is conceptual (as described above).
This is accomplished by generating a thermal shock. That is, as a result of the temperature change, the cells detach and retain their original shape as a monolayer on the slide. To understand how this method works, think of what it feels like, for example, when you touch a frozen surface with your bare hands.

In other words, if you touch a frozen surface with your bare hands, you will immediately feel that your hands are glued to the surface. In fact, when a frozen surface is touched, a thin layer of cells is left behind on the frozen surface.

That is, the present invention includes the following steps: (A) Slide the cell section sample into two culms, () l) Rapidly cool the slide to apply a thermal shock to the cell tissue, and (0 ) A method for preparing a cell tissue specimen, which comprises the step of removing a cell section sample from a slide, and an apparatus used therefor.

The apparatus of the present invention is composed of at least the following (a) to (0).

(a) A cryocooler capable of sprinkling a cryogenic liquid onto the surface of a slide, thereby making it possible to bring the surface below -4°.

(b) at least one fan used to promote the volatilization of the cryogen-generating liquid and to generate the required cold temperature on the slide;

(c) The temperature can be adjusted to a predetermined temperature using a thermostat, and when the slide reaches the ideal temperature, an automatic system that applies pressure to the sectioned sample of the organ under test that is inserted between the apositor and the slide. apositor.

In addition to the above (a) to (a), (d) it is possible to provide an automatic slide mover that makes it possible to apply different pressures on the same slide. .

FIG. 1 shows a partially cutaway side view of the cryopreposition device of the present invention. In FIG. 1, (1) is a storage cylinder for a low temperature generation liquid, and (2) is a low temperature generation liquid (for example, ethyl chloride). (3) is above the liquid for low temperature generation, and is a slightly high pressure gas. (4) is a cock, and by opening it, the liquid for low temperature generation is released.
2) some of the circuit (5) and nozzle (6)
Through this, it becomes possible to sprinkle the liquid onto a limited portion of the tube of the slide (9). (7) and (8) are fans, and the jet stream generated by them is controlled by deflectors (7) and (8) and directed toward the bottom of the slide (9) to evaporate the low temperature generating liquid. promote. In this way the cooling of the slide (9) is made more rapid. The slide (9) is controlled so as to be able to move laterally by suitable guides ◇0) and (11). Detachment (The section of the organ to be offered is placed on a slide (9) to be compressed by an apositor (b).
This pressure can be gradually applied using the toothed shirt (18) and the pinion gear θ.

In addition, when pressing the organ section sample, the slide (9) is kept at a constant temperature of G.
It will be carried out after reaching this point. It usually takes 6 to 4 minutes after the cock (4) is opened and the fans (7) and (8) are turned on almost simultaneously for the temperature to become constant. Finally, the guides 00) and (11) are vibrated by the vibrator (15) (so that the slide is vibrated at the same time). This vibration makes it easy to peel off the organ sample sections that were adhered by thermal shock.

Furthermore, this device allows the slide (9) to be automatically moved by the guides (a) and (11), so that the pressure when compressing is transmitted through the slide and adheres to the component parts of the slide. (apposition). Therefore, the first attachment part, the second attachment part
It is possible to obtain information about cell layers that have different degrees of peeling, such as the appended part, the sixth attached part, and the fourth added part. In this way, cell properties that are indistinguishable by normal histological testing methods can be restored layer by layer. The degree of adhesion of cells to the cooled slides can also be estimated by the index C of neoplasia θ density.

The mounting W shown in the drawing (FIG. 1) and the description above can be modified as desired within the scope of the present invention. For example, the fans (7) and (8) and the vibrator (15) may be electrically operated by batteries or electric current. The low temperature generation liquid may be replaced by a gaseous substance (for example, liquefied nitrogen) G that is in a liquid state at a low temperature, if the special effect of the low temperature is compromised. The pressurized gas (8) may be the vapor of a liquid for low temperature generation. In that case, for example, a sleeve having a heating effect may be provided around the cylinder (1), and when the cock (4) is opened. At the same time, the cylinder (1) is heated to gasify the liquid (2), and the gas (8) is brought to a higher pressure and sent to the nozzle (6).
The characteristics of the deposition device of the present invention are that it is small (typically 60 to 4 Qcm in size) and very easy to handle. The main reason for this is that it is compact and very easy to transport.

The cooling time varies depending on the slant of the section sample and the thickness of the slide, but in any case, the cooling time is very short, usually several tens of seconds. At the end of the cooling period, the section specimen is removed with tweezers and the remaining slide (9) is placed in the guide (10).
and (11) Colored immediately after removal from the bottle.

Additionally, slides may be prestained, for example, if the time required from the time the instrument is started to the time the slides are submitted for diagnostic analysis must be less than 6 to 5 minutes. You can also do it.

When a cell tissue specimen is prepared using the apparatus of the present invention and a pathological test for diagnosis is performed, extremely important information can be obtained in a short period of time.

(b) Structure of the parenchyma (parθnchyrna) subjected to the test, peeled in a single layer onto a slide.

(b) Parenchyma due to the difference in the density of the stripped cells (
par+Jnobyoa) property difference X.

e-) On the negative side, the presence of vascular structures.

Every time.

Additionally, the low humidity deposition device of the present invention allows for highly detailed cellular diagnostic testing.

For example, the nucleus (nuc, teus), micronucleus (nuo
leoli), mitosis, mast cells (mast c
θ11) and their quantitative ratio between cells and abnormal cells. 1!1 for abnormal cells! These include neo-acidoid cells caused by tumors, cells with immune reactivity, and pathological tissues caused by hepatitis, and each can be examined in detail. Furthermore, the inflammatory response in the tumor area can also be investigated.

[Brief explanation of drawings]

Figure jA1 is a partially cutaway side view of the cryoadhesion device of the present invention. (Main symbols in the drawing) (1): Liquid storage cylinder (2): Liquid for low temperature generation (3): High pressure gas (7) and (8): 7 Ann (9) Nisuride (B) Near positor (15): vibrator

Claims (1)

[Claims] 1(A) Step of placing the cell section sample on a slide (
B) A method for preparing a cell tissue specimen comprising the steps of rapidly cooling the slide to apply a thermal shock to the cell section sample, and (0) removing the cell section sample from the slide. 2. The method according to claim 1, wherein the cooling of the slide in (B) is cooling to at least −40° or less. 3. The method according to claim 1, wherein in the step (B), the thermally shocked cell slice sample is compressed. 4. The method according to claim 1, wherein in step (0), the operation of removing the cell slice sample from the slide is performed by mechanical vibration of the slide. 5 (a) Freezer for spraying a low temperature generating liquid onto the slide surface that can bring the surface of the slide below -4°C; (b) Accelerating the volatilization of the sprayed low temperature generating liquid to increase the temperature of the slide surface. at least one fan for efficient lowering; and (0) applying a measurable and gradually increasing pressure to the specimen section placed between the slide and the apositor when the slide reaches a certain temperature. A cell tissue specimen preparation device consisting of an automatic apositor that applies voltage. 6. The device according to claim 5, further comprising means for adjusting the temperature of the apositor. 7. Apparatus according to claim 5, further comprising means for automatic movement of the slide in order to deposit a number of cell tissues on one slide. 8. The device according to claim 7, further comprising means for varying the applied pressure as the slide moves. 9. The apparatus of claim 5, wherein the freezer has a cylinder for storing a cryogenic liquid. 10 Claim 5, wherein the freezer has a nozzle capable of spraying a low temperature generating liquid onto the center of the slide.
The device according to paragraph 9 or paragraph 9. 11. The device according to claim 5, wherein the low humidity generating liquid is a low boiling point liquid. 12. The apparatus of claim 11, wherein the low boiling point liquid is ethyl chloride. 13. The apparatus according to claim 5, wherein the low temperature generating liquid is a liquefied gas. 14. The device according to item 16, wherein the liquefied gas is liquefied nitrogen. 15 Claim t+I4 in which means are added for increasing the rate of discharge of the cryogenic liquid from the cylinder
16. The device according to claim 9, wherein the means is the presence of a high pressure gas above the liquid in the cylinder. The device according to claim 15. 17. The device according to claim 15, wherein the means is to heat the cylinder. Scissors iM according to item 15.