CN1205078A - Liquid treatment method for samples for microscopy - Google Patents

Abstract

Liquid treatment method for samples for microscopy. An apparatus for automatically dyeing samples for microscopy, wherein slide glasses (12) supported on a dyeing cage (6) are immersed completely in the cleaning water (11) stored in a liquid storage vessel (2), and thereafter pull up the slide glasses completely from the cleaning water (11). These actions are made repeatedly to promote the replacement of the cleaning water in the clearances (13) between adjacent slide glasses (12) by new cleaning water. Thus, an operation for washing away the chemicals deposited on the slide glasses (12) can be carried out effectively for conducting a sample dyeing treatment.

Description

Liquid handling methods for specimens for microscopic observation

technical field

The present invention relates to a method of liquid treatment of a piece of biological tissue or cell (hereinafter referred to as a sample) to be observed with a microscope in a clinic, hospital or various research institutes, particularly a method for microscopic observation using staining treatment. In the automatic staining device of specimens, the improvement of the method of washing off the liquid medicine attached to the glass slide.

Background technique

In clinics and hospitals, a sample excised from an affected area is usually observed under a microscope for diagnosis of a disease. In order to easily observe and diagnose diseases with a microscope, a sample attached to a glass slide is stained, and various automatic staining devices for automatically performing such staining work have been provided.

FIG. 2 shows an example of such an automatic dyeing apparatus, which is described in Japanese Patent Laid-Open No. 5-66543. In the upper part of the cabinet 1, a plurality of liquid storage containers 2 with upper openings and storing various chemical liquids necessary for the dyeing process are arranged in a row. A part of these liquid storage containers 2 are washing containers in which washing water such as tap water can be freely supplied, and the rest are chemical liquid containers storing various chemical liquids necessary for dyeing work. The top of these liquid storage containers 2 is provided with a three-dimensional driving device, by the movable beam 3 that can move freely horizontally in the transverse direction (a direction of Figure 2) of the casing 1, by moving freely along the movable beam 3, it can move along the horizontal direction of the casing 1. A moving column 4 that can move horizontally in the longitudinal direction (direction b in FIG. 2 ) and a cantilever 5 that moves vertically in the vertical direction of the moving column 4 can be freely raised and lowered.

As shown in Figure 3, the dyeing cage 6 on the cantilever 5 can be freely loaded and unloaded. That is, the staining cage 6 has a slide receiving portion 7 and a handle 8 , and a bonding piece 9 is provided at the center of the handle 8 . The front end of the cantilever 5 is freely engaged with the engagement piece 9 by being inserted into the engagement fixing hole 10 provided on the engagement piece 9 . Then, by lowering the cantilever 5 in the state where the cantilever 5 is engaged with the joint piece 9 , the dyeing cage 6 is freely immersed in the liquid storage container 2 .

When staining a sample with the above-mentioned automatic staining apparatus, first, the staining cage 6 is joined to the cantilever 5, and the staining cage 6 accommodates a slide glass on which a sample to be stained is attached. Next, the above-mentioned three-dimensional driving device is driven to move the moving beam 3, moving column 4, and boom 5 constituting the three-dimensional driving device according to signals from a controller not shown in the figure. Then, the above-mentioned dyeing cage 6 is immersed in the liquid storage container 2 at a predetermined time according to a predetermined procedure. By repeating such an operation a predetermined number of times, the sample is dyed. In addition, using the above-mentioned automatic staining device, a plurality of staining cages 6 are processed side by side to efficiently stain more samples. The multiple staining process has also been described in Japanese Patent Publication No. 6-100522, which is known in the art.

That is, the staining cage 6 containing the slide glass on which the sample is attached is immersed in the chemical solution for a predetermined period of time. During the immersion, the cantilever 5 is disengaged from the staining cage 6 and engages with other staining cages 6. And other dyeing cages 6 are sent in the liquid storage container 2 of regulation. Similarly, the connection with the cantilever 5 is disengaged, and the other dyeing cage 6 is immersed in the chemical solution only for a predetermined time. The above-mentioned bonding is performed on a plurality of dyeing cages 6 in parallel, and if there is a dyeing cage 6 that has been dipped for a predetermined time, the cantilever 5 is bonded to the dyeing cage 6 and moved to the next liquid storage container 2 . By repeating such operations, since a plurality of staining cages 6 can be processed in parallel, more samples can be stained in a short time compared with the above-mentioned normal processing. Such multiple staining processing can be realized by storing a processing program in advance in the above-mentioned controller.

When staining the sample attached to the slide glass as described above, after the above-mentioned staining cage 6 is extracted from the first liquid medicine stored in the predetermined liquid storage container 2, the staining cage 6 is immersed in another Before adding the second medicinal solution stored in the liquid storage container 2, the first medicinal solution adhering to the staining cage 6 and the slide glass in the staining cage 6 must be washed away. The reason is to stop the chemical reaction of the first chemical liquid, or prevent the first chemical liquid from being mixed into the second chemical liquid to deteriorate the second chemical liquid.

Thus, in the prior art, as shown in FIGS. 4A and 4B , after the above-mentioned staining cage 6 is extracted from the first liquid medicine, the staining cage 6 and the stained cage 6 attached to the staining cage 6 on the slide 12 are washed away. A liquid medicine. That is, after the cantilever 5 on which the dyeing cage 6 is suspended is moved above the liquid storage container (washing container) 2 storing the washing water 11, the cantilever 5 is lowered to immerse the dyeing cage 6 in the washing water 11. Then, by lifting and lowering the cantilever 5 slightly, the above-mentioned staining cage 6 is displaced up and down between the state of FIG. 4A and the state of FIG. Attached first liquid medicine.

In the conventional case, the lifting and lowering of the staining cage 6 in the washing water 11 is carried out under the following conditions, that is, all or a part of the sample adhering to the surface of the slide glass 12 accommodated in the staining cage 6 is immersed in the 11 in the above cleaning water. As a result, among the cleaning water 11 entering the gaps 13 between the samples on the surfaces of the adjacent slide glasses 12, it is difficult to replace the old cleaning water existing in the lower part of the gaps 13 with new cleaning water. As a result, among the first chemical solutions adhering to the respective slide glasses 12 , the first chemical solutions adhering to the portions facing the lower portions of the respective gaps 13 are difficult to wash off.

Therefore, the previous washing operation required a long time, that is, several times of lifting and lowering was required to wash away the above-mentioned first liquid medicine. Therefore, the time required for the dyeing work becomes longer. Furthermore, when a plurality of dyeing cages 6 are processed in parallel at the same time, if the time for using the cantilever 5 is prolonged due to the washing operation, the processing of other dyeing cages 6 cannot be performed.

The object of the present invention is to solve the above-mentioned problems, and to provide a liquid treatment method for a specimen for microscopic observation which can more effectively wash away the chemical liquid adhering to a slide glass than conventionally.

disclosure of invention

A liquid processing method provided by the present invention is to sequentially immerse the cage into the liquid in a plurality of liquid storage containers to process the sample, wherein a plurality of glass slides to be used as specimen samples for microscope observation are affixed to correspond to each other. There are gaps between adjacent slides, and they are stored in the cage in an upright manner. The method includes relatively lowering the cage to a plane where the upper edge of the slides is below the liquid surface in the liquid storage container. The step of immersing in the liquid, and then, the step of raising the cage relative to the surface of the slide glass until the lower edge of the sample on the surface of the slide glass is located above the plane above the liquid surface in the liquid storage container. Usually, the aforementioned steps are repeated.

The present invention also provides a liquid treatment method, in which the cage is sequentially dipped into the liquid in a plurality of liquid storage containers including the liquid medicine container and the cleaning liquid container to process the sample, wherein the specimen sample to be used as a microscope observation is attached. A plurality of slides are housed in the cage in such a manner that there are gaps between adjacent slides and stand upright in the vertical direction, the method includes immersing the cage in the liquid medicine in the liquid medicine container, A step of treating the sample on the slide glass with a chemical solution, and then, relatively lowering the cage in the cleaning solution in the cleaning solution container to a plane where the upper end edge of the slide glass is located below the surface of the cleaning solution so as to be immersed in the The step of washing in the liquid, and then, the step of raising the cage relative to the surface of the glass slide until the lower edge of the sample is located above the plane above the plane of the washing liquid. Usually, the step of immersing in the cleaning liquid and the step of rising to the surface of the cleaning liquid are repeated.

The aforementioned steps repeated can be carried out in the same cleaning solution container, or can be carried out in multiple different cleaning solution containers. The step of treating with the aforementioned chemical solution is a typical dyeing step. In addition, the aforementioned washing liquid is typically water.

In the present invention, by employing the above-mentioned steps, most of the cleaning solution containing the chemical solution to be washed away existing in the gap between adjacent slide glasses is discharged downward from the gap. Therefore, in the state where the slide glass is immersed in the cleaning solution again, the chemical solution to be washed away or the new cleaning solution with a small amount of the chemical solution mixed therein does not enter the gap. As a result, it is possible to efficiently wash away the chemical solution adhering to the portion of the slide glass that faces the gap.

A brief description of the graphics

FIGS. 1A and 1B schematically show longitudinal sectional views of a state in which a chemical solution adhering to a slide glass is washed away with a cleaning solution by the liquid treatment method of the present invention.

Fig. 2 is a perspective view showing an example of an automatic staining device for a specimen for microscopic observation for carrying out the liquid processing method of the present invention.

Figure 3 shows a perspective view of an example of a staining cage.

4A and 4B schematically show longitudinal cross-sectional views of a state in which a chemical solution adhering to a slide glass is washed away with washing water by a conventional method.

Best form for carrying out the invention

Figure 1 shows an embodiment of the invention. The liquid processing method of the present invention is characterized in that, in the automatic staining device for specimens for microscopic observation in a known form as described in FIG. Before storing other liquid medicines in the liquid medicine container, the above-mentioned prescribed liquid medicines adhering to the staining cage 6 and the slide glass 12 accommodated in the staining cage 6 can be effectively washed away. The structure and function of the automatic staining device for specimens for microscopic observation are not limited to the conventional structure shown in FIG. 2. Therefore, for the sake of omitting description, only the characteristic parts of the present invention will be described below.

Before the staining cage 6 lifted from the chemical solution container storing the above-mentioned predetermined chemical solution enters the chemical solution container storing other chemical solutions, in order to wash away the slide glass 12 on the staining cage 6 and accommodated in the staining cage 6 The above-mentioned prescribed medicinal liquid attached on the top is moved to the top of the liquid storage container 2 as a washing container for storing the cleaning water 11 by hanging the cantilever 5 of the dyeing cage 6 . Next, the cantilever 5 is lowered, and the staining cage 6 suspended by the cantilever 5 is immersed in the cleaning water 11 stored in the above-mentioned liquid storage container 2 as shown in FIG. 1A until the slide glass 12 accommodated in the staining cage 6 until the upper edge is below the water level. Thereafter, the cantilever 5 is raised so that, as shown in FIG. 1B , the lower edge of each slide glass 12 is positioned on a horizontal plane. At this time, part of the lower edge of the slide glass 12 may be below the horizontal plane, but the sample itself on the surface of the slide glass may be entirely on the horizontal plane. Next, the state of FIG. 1A and the state of FIG. 1B are repeated a predetermined number of times by lowering the boom 5 .

Adopt above-mentioned structure, when carrying out the liquid processing method of specimen for microscopic observation of the present invention, as shown in Figure 1B, when the cantilever 5 is raised to be accommodated in the slide glass 12 surface in the staining cage 6, the lower end edge of the specimen is positioned In the state above the plane on the horizontal plane, the chemical solution to be washed away exists in the gap 13 between adjacent slides 12 or between the surfaces of slides adjacent to the sample on the surface of the slides. Most of the cleaning water is discharged from the gap 13 to below under the action of gravity. In order to promote the downward discharge of the cleaning water mixed with the chemical solution from the gap 13, the staining cage 6 and the slide glass 12 inside it may also be vibrated. Then, while the slide glass 12 is immersed in the cleaning water 11 again, new cleaning water with a small amount of the chemical solution to be washed off flows into the gap 13 . As a result, the chemical solution adhering to the part of the slide glass 12 that faces the gap 13 can be effectively washed away.

In addition, in the illustrated example, the operation of raising the cantilever 5 after dipping the dyeing cage 6 in the washing water 11 is performed in the same liquid storage container 2 . Corresponding thereto, as a water washing container, a plurality of liquid storage containers 2 storing cleaning water 11 can be used. After the above-mentioned dyeing cage 6 is immersed in the cleaning water 11 in the first liquid storage container, the Bring out the dyeing cage 6 and immerse the dyeing cage 6 in the cleaning water 11 in the second, third... liquid storage container. In addition, the dyeing cage 6 and the liquid storage container 2 can be raised and lowered relative to each other, or the upper and lower positions of the dyeing cage 6 can be kept still while the liquid storage container 2 can be raised and lowered. But, preferably, make dyeing cage 6 lift, the energy required for lifting like this is less, and the liquid in the liquid storage container 2 is difficult for overflowing.

The liquid processing method of the sample for microscopic observation of the present invention can effectively wash away the chemical liquid attached to the slide glass due to the above-mentioned structure, and can shorten the time required for sample processing such as staining processing. In addition, when multiple staining cages are processed in parallel, the time to use the cantilever for cleaning is shortened, making it easier to process other staining cages.

By using the liquid treatment method of the present invention, as a result of effectively washing away the liquid medicine, there is no sample treatment unevenness, which is a typical phenomenon of uneven dyeing, and the dyeing performance is improved. Moreover, during staining, the remaining staining solution attached to the slide and staining cage can be washed away with a small amount of cleaning solution in a short time. In addition, during cleaning and when the outside is dipped into the liquid storage container during cleaning, the number of stirring actions of the slide cage can be reduced, and this effect is more remarkable when multiple slide cages are processed in parallel. Conventionally, when the slide cage is immersed in the cleaning solution container and the chemical solution container, the cleaning solution and the chemical solution are continuously stirred, but it is also very advantageous to reduce the frequency of stirring. In addition, the liquid handling method of the present invention can effectively replace the liquid between the slides, and as a result, the distance between the slides can be reduced, and the arrangement length of the slides can be reduced.

Possibility of industrial use

The liquid processing method of the present invention is not limited to attaching a sample such as a biological tissue piece or a cell on the surface of a glass slide, and washing the slide glass supported by the staining cage after staining, and it can also be used in a place other than the washing container. In the case of a reservoir, displaces the liquid in the gap between the slides.

Claims (8)

1, a kind of method for treating liquids of samples for microscopy, be that cage is impregnated in the liquid in a plurality of storage liquid containers successively to handle sample, wherein post and to be contained in the described cage to have gap, the upright mode of above-below direction between adjacent microslide as a plurality of microslides of the sample of samples for microscopy, this method comprises

The last ora terminalis that described cage is dropped to microslide relatively is arranged in till the subsurface plane of storage liquid container to be immersed in the step of liquid, then,

Described cage is risen to step till the plane more than the plane on the liquid level that ora terminalis under the sample on the slide surface is positioned at the storage liquid container relatively.

2, according to the described method for treating liquids of claim 1, it is characterized in that, carry out described step repeatedly.

3, a kind of method for treating liquids of samples for microscopy, cage is impregnated into successively in the liquid in a plurality of storage liquid containers that comprise medical liquid container and soda liquor container to handle sample, wherein post and to be contained in the described cage to have gap, the upright mode of above-below direction between adjacent microslide as a plurality of microslides of the sample of samples for microscopy, this method comprises

Described cage is immersed in the soup in the described medical liquid container, handles the step of sample on the microslide with soup, then,

With drop to relatively in the cleaning fluid of described cage in described soda liquor container microslide on ora terminalis be arranged in and clean till the subsurface plane to be immersed in the step of cleaning fluid, afterwards,

Described cage is risen to step till ora terminalis under the sample on the slide surface is positioned at plane more than the plane of cleaning on the liquid level relatively.

4, according to the described method for treating liquids of claim 3, it is characterized in that, be impregnated into the step and the step that rises on described cleaning liquid level of described cleaning fluid repeatedly.

According to the described method for treating liquids of claim 4, it is characterized in that 5, the described step of carrying out is repeatedly carried out with same soda liquor container.

According to the described method for treating liquids of claim 4, it is characterized in that 6, the described step of carrying out is repeatedly carried out with different a plurality of soda liquor containers.

According to the described method for treating liquids of claim 3, it is characterized in that 7, the step of handling with described soup is a staining procedure.

According to the described method for treating liquids of claim 3, it is characterized in that 8, described cleaning fluid is a water.

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