JPH04268435A - Method and apparatus for stretching chromosome on slide glass - Google Patents

Abstract

PURPOSE:To easily stretch a chromosome onto an upper surface of a slide glass. CONSTITUTION:A slide glass 12 is taken out of a cooling chamber 6, and the surface is clouded up with steam in a high temperature and highly humid clouding chamber 7. A nucleus is dripped down on the upper surface of the slide glass 12 with its surface clouded with steam in a stretching chamber 8 and stretched on the surface of the slide glass 12. Then while it passes through a first drying chamber 9 and a second drying chamber 10, the surface of the slide glass 12 is dried, and then the slide glass 12 is taken out.

Description

[Detailed description of the invention]

0001

[Industrial Application Field] The method and apparatus for spreading chromosomes on a slide glass according to the present invention are useful for diagnosing diseases such as leukemia and various malignant tumors, and determining genetic disease status. Used to spread (stretch) genes onto a glass slide.

0002

[Prior Art and its Problems] In order to diagnose diseases such as leukemia and various malignant tumors, and to determine genetic disease status, chromosomes of patients, etc. are sometimes examined.

[0003] When performing such a chromosome test, the nucleus in the cell collected from the patient or parent is taken out, and the chromosomes 1 and 1 in this nucleus are spread out on a glass slide as shown in Figure 7. Chromosomes 1 and 1 are then observed using a microscope.

[0004] As described above, when chromosomes 1 and 1 are spread on the top surface of a slide glass, they must be spread sufficiently. This is because, as shown in Figure 8, if chromosomes 1 and 1 overlap one another, it is not possible to sufficiently observe the overlapping chromosomes 1 and 1, as shown in Figure 9.
This is because it is necessary to shift each chromosome 1, 1 in the horizontal direction.

[0005] Conventionally, methods such as flame drying, air drying, and steam drying have been known as methods for spreading the chromosomes in the cell nucleus onto a glass slide so that the chromosomes 1 and 1 do not overlap. There is.

[0006] However, all of the conventional chromosome extension methods must be carried out manually by the operator and require skill, making the work of preparing chromosome specimens troublesome and requiring the processing of a large number of specimens in a short period of time. It was difficult to do.

[0007] The method and apparatus for spreading chromosomes on a slide glass according to the present invention were devised in view of these circumstances.

[0008]

[Means for Solving the Problem] Among the method and apparatus for spreading chromosomes on a slide glass of the present invention, the invention of the spreading method described in claim 1 is characterized in that a cooled slide glass is placed in a high temperature and high humidity atmosphere. After moistening the surface of the slide glass by exposing it to water, Carnoy's fixative containing cell nuclei is dropped onto the top surface of the slide glass to spread the chromosomes in the cell nuclei on the top surface of the slide glass, and then the slide glass is exposed to low humidity. The sample is transferred to an atmosphere to evaporate the water and Carnoy's fixative on the top surface of the slide glass, leaving only the chromosomes in an extended state on the top surface of the slide glass.

[0009] Furthermore, the invention of the stretching device described in claim 2 includes a cooling chamber for storing a plurality of glass slides in a cooled state, and a transporting device for transporting the glass slides taken out one by one from the cooling chamber. A stretching chamber provided in the middle of the transfer means and having a high temperature and high humidity inside, and a drying chamber provided in the middle of the transfer means and rearward of the stretching chamber in the transfer direction.

0010

[Operation] In the method and apparatus for spreading chromosomes on a slide glass of the present invention, which is constructed as described above, the chromosomes in the cell nucleus that have been dropped together with Carnoy's fixative on the top surface of the slide glass are taken out, and the chromosomes are further spread. After spreading it on the top surface of the slide glass, dry the top surface of the slide glass,
It is possible to automatically leave only the chromosomes on the top of the slide glass.

[0011]

DESCRIPTION OF THE PREFERRED EMBODIMENTS Next, the present invention will be explained in more detail with reference to the illustrated embodiments. 1 to 6 show an embodiment of the apparatus for spreading chromosomes on a slide glass according to the present invention, FIG. 1 is a perspective view showing the overall configuration, and FIG. 2 is an enlarged A--
A sectional view, FIG. 3 is a BB sectional view in FIG. 4, and FIG. 4 is a C in FIG. 2.
-C sectional view, FIG. 5 is a perspective view seen from above in FIG. 4 with the cover removed, and FIG. 6 is a DD sectional view in FIG. 2.

An extension unit 3 is provided on the front side of the top surface of the substrate 2, and a control unit 4 is provided on the back side of the top surface. The stretching unit 3 includes a cooling chamber 6, a fogging chamber 7, and a stretching chamber 8, arranged in order from the front side in the feeding direction on the upper surface of a swinging plate 5 provided on the upper surface of the substrate 2 at a freely tiltable angle. It is constructed by arranging a first drying chamber 9 and a second drying chamber 10 in a straight line.

That is, one end edge of the swing plate 5 having the extension unit 3 on its upper surface is pivotally supported on the upper surface of the base plate 2 by a hinge 30. Further, the studs 31 fixed to the opposite ends of the swing plate 5 are loosely engaged with the long holes 33 of the support plate 32 fixed to the upper surface of the substrate 2, as shown in FIG. A nut 34 is screwed onto the end of the stud 31, and by tightening the nut 34, the stud 31 can be fixed at a desired position in the elongated hole 33. By adjusting the fixing position of the stud 31, the inclination angle of the swing plate 5 provided with the extension unit 3 changes within a range of 5 to 10 degrees.

Reference numeral 11 denotes a chain conveyor which is a transfer means for transferring the slide glasses 12, and the slide glasses 12 taken out one by one from the cooling chamber 6 are transferred to the fogging chamber 7, the stretching chamber 8, and the first chamber. Drying chamber 9 and second drying chamber 1
0 and then delivered to the end of the extension unit 3.

Among the above-mentioned chambers 6 to 10, first, inside the cooling chamber 6, a guide rail 13 extending in the vertical direction is provided. Slide glasses 12 are held at intervals in the vertical direction. Then, the slide glasses 12 held in the holder 14 are removed one by one from the chain conveyor 1 by a conventionally known take-out means.
1 and can be sent out freely.

In FIG. 1, reference numeral 15 denotes a cooling unit, which uses a built-in cooler to generate cold air, which can be freely supplied into the cooling chamber 6. That is, the cooling unit 15 and the cooling chamber 6
are connected by a pair of hoses 16, 16, and the air sucked from the cooling chamber 6 into the cooling unit 15 through one hose 16 is cooled and then returned to the cooling chamber 6 again. Therefore, the inside of the cooling chamber 6 is kept at a low temperature.

A heat exchanger 18 for humidification is provided in the fog adhesion chamber 7 adjacent to the cooling chamber 6 via the partition wall 17, as shown in FIGS. Then, the other end of the hose 19, one end of which was connected to the heat exchanger 18, is connected to the humidifier 20.
is connected to. Water 2 stored at the bottom of this humidifier 20
A porous nozzle 22 is sunk in 1, and the air discharged from the compressor 23 is passed through the porous nozzle 22.
The structure is such that the water is ejected into the water 21 through the water. Therefore, the relative humidity in the space above the humidifier 20 is approximately 1.
00% air is produced.

In this way, the humid air generated in the upper space of the humidifier 20 is pushed into the hose 19 by the air jetted from the porous nozzle 22, and the air is pushed out into the hose 19, and the air is forced out into the hose 19.
Sent within 8. Then, the air is heated while passing through the heat exchanger 18, which has been set to a desired temperature by a heat source such as the thermo module 40, and becomes hot and humid air having the desired temperature and humidity. Blowing out inside.

On the other hand, a cooling plate 24 is provided at the bottom of the fogging chamber 7 at a position above which the slide glass 12 sent by the chain conveyor 11 passes, in order to keep the slide glass 12 at a low temperature. ing. This cooling plate 24 is formed with drain grooves 27, 27, so that condensed water generated on the surface of the cooling plate 24 can be freely discharged. and,
The condensed water discharged from the cooling plate is discharged from a drain port 35 provided at the end of the fogging chamber 7.

Reference numerals 25 and 25 denote guide rails disposed above the cooling plate 24. Both ends of the slide glass 12 are supported by the guide rails 25 and 25, and the pressing pieces provided on the chain conveyor 11 are supported at both ends of the slide glass 12. 26, 26 (Fig.
) and sent from the cooling chamber 6 to the second drying chamber 10.

Furthermore, a stretching chamber 8 and first and second drying chambers 9,
Also inside the chambers 10, as shown in FIG. 6, heat exchangers 29 each connected to a humidifier 28 are provided so that the temperature and humidity in each chamber 8 to 10 can be adjusted to desired values. There is. However, the inside of the stretching chamber 8 is kept in a high humidity state, the inside of the first drying room 9 is kept in a slightly low humidity state, and the inside of the second drying room 10 is kept in an even lower humidity state. The humidity in the stretching chamber 8 and each drying chamber 9, 10 is controlled by a humidifier 2.
0, 28 and the heat exchangers 18, 29. That is, the lower the temperature of the humidifiers 20, 28, or the higher the temperature of the heat exchangers 18, 29, the lower the humidity in the stretching chamber 8 and each drying chamber 9, 10.

[0022] A control panel 36 is provided on the front of the control unit 4, and inside the humidifier 2, in addition to the controls for each part, is provided.
0 and 28 are provided. Also, the lower edge of the partition wall 17 that partitions the cooling chamber 6 and the fogging chamber 7, the lower edge of the partition wall 37 that partitions the fogging chamber 7 and the extension chamber 8, and the partition wall that partitions the extension chamber 8 and the first drying chamber 9. A shutter 39 that opens only when the slide glass 12 passes is provided at the lower edge of each of the shutters 38 .

When the apparatus for spreading chromosomes on a slide glass of the present invention constructed as described above is used to spread chromosomes on the upper surface of the slide glass 2, the procedure is as follows.

First, only one slide glass 12 is taken out from the holder 14 in the cooling chamber 6 and sent to the fogging chamber 7. Because the inside of the fogging chamber 7 is hot and humid, when a cooled slide glass 12 is sent into the fogging chamber 7, the water vapor that comes into contact with the slide glass 12 condenses and moistens the surface of the slide glass 12. let

The slide glass 12 whose surface has been moistened in this manner is then fed into the extension chamber 8. Then, a small amount of Carnoy's fixative containing cell nuclei is dropped onto the upper surface of the slide glass 12 using a dropping nozzle (not shown). methanol and glacial acetic acid in a ratio of 3:1 (methanol:
Since Carnoy's fixative mixed with glacial acetic acid = 3:1 mixes with water in any ratio, the Carnoy's fixative containing metaphase cell nuclei dropped onto the top surface of the slide glass 12, which is wet over the entire surface, is , spread over the top surface of this slide glass 12.

At the same time, the cell membrane surrounding the cell nucleus ruptures, and the chromosomes in the cell nucleus spread (extend) on the top surface of the slide glass 12 together with Carnoy's fixative. Particularly in the case of this embodiment, since the entire stretching unit 3, including the chain conveyor 11 for transporting the slide glass 12, is supported in an inclined state, the chromosomes can be stretched efficiently and reliably. However, depending on the case, the slide glass 12 may remain horizontal.

[0027] It has been confirmed through experiments by the present inventors that the cell membrane ruptures and the chromosomes expand in this way, and this is thought to be due to the following reasons.

That is, when a cell nucleus is dropped onto the top surface of a moist slide glass 12, water enters the cell nucleus due to the osmotic pressure, the cell membrane surrounding the cell nucleus swells, and finally this cell membrane ruptures, causing the inside of the cell nucleus to swell. It is thought that the chromosomes are spread on the top surface of the slide glass 12 together with Carnoy's fixative.

The slide glass 12 with the chromosomes spread on its upper surface in this manner is sent by the chain conveyor 11,
It is sent into the first drying chamber 9.

While passing through the first drying chamber 9 kept at relatively low humidity, the surface of the slide glass 1 has been slightly dried.
2 is a second drying chamber 1 which is then maintained at a lower humidity.
The water adhering to the upper surface of the slide glass 12 and the Carnoy's fixative solution dropped onto the upper surface are almost completely evaporated. As a result, the slide glass 12
Only the chromosomes remain on the top surface.

Therefore, the slide glass 12 with chromosomes attached in an extended state on its upper surface is sent out to the outside of the second drying chamber 10 and moved to the next step, where a cover glass is placed on the upper surface of this slide glass 12. , and attach it so as to cover the chromosomes mentioned above.

[0032] These series of operations can be easily performed without requiring special skill, and furthermore, the stretching operation can be automated.

[0033]

[Effects of the Invention] The method and device for spreading chromosomes on a slide glass of the present invention are constructed and operated as described above, and therefore, the chromosomes in the cell nucleus can be spread on the top surface of the slide glass without any troublesome work. It is possible to automate the chromosome extension work and efficiently process a large number of specimens.

[Brief explanation of the drawing]

FIG. 1 is a perspective view showing the overall configuration of an embodiment.

FIG. 2 is an enlarged cross-sectional view taken along line AA in FIG. 1;

FIG. 3 is a cross-sectional view taken along line BB in FIG. 4;

FIG. 4 is a sectional view taken along line CC in FIG. 2;

FIG. 5 is a perspective view from above of FIG. 4 with the cover removed.

[Figure 6] DD sectional view in Figure 2

FIG. 7 is an enlarged plan view showing chromosomes spread out on the top surface of a slide glass.

FIG. 8 is an enlarged longitudinal cross-sectional view showing a state in which the chromosomes are not fully extended and overlap.

FIG. 9 is an enlarged longitudinal cross-sectional view showing a state in which the chromosomes are well expanded.

[Explanation of sign]

1 Chromosome 2 Board 3 Extension unit 4 Control unit 5 Rocking plate 6 Cooling room 7. Cloudy adhesion chamber 8 Extension room 9 First drying room 10 Second drying room 11 Chain conveyor 12 Slide glass 13 Guide rail 14 Holder 15 Cooling unit 16 Hose 17 Partition wall 18 Heat exchanger 19 Hose 20 Humidifier 21 Water 22 Porous nozzle 23 Compressor 24 Cooling plate 25 Guide rail 26 Pressing piece 27 Drainage ditch 28 Humidifier 29 Heat exchanger 30 Hinge 31 Stud 32 Support plate 33 Long hole 34 Nut 35 Drain port 36 Control panel 37 Partition wall 38 Partition wall 39 Shutter 40 Thermo module

Claims (2)

[Claims] Claim 1: After moistening the surface of a cooled slide glass by exposing it to a high-temperature, high-humidity atmosphere, Carnoy's fixative containing cell nuclei is dripped onto the top surface of the slide glass, and the chromosomes in the cell nuclei are After spreading the above slide glass on the top surface of the slide glass, the slide glass is transferred to a low humidity atmosphere to evaporate the water and Carnoy's fixative on the top surface of the slide glass, leaving only the chromosomes in a stretched state on the top surface of the slide glass. A method of spreading chromosomes on glass. 2. A cooling chamber for storing a plurality of glass slides in a cooled state; a transfer means for transferring the slide glasses taken out one by one from the cooling chamber; An apparatus for spreading chromosomes on a slide glass, comprising an extension chamber kept at high temperature and high humidity, and a drying chamber provided midway through the transfer means and rearward of the extension chamber in the transfer direction.