JPS58196438A - Method and device for dyeing biological sample on microscope slide - 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 (8) The present invention relates generally to the microscopic examination of biological specimens mounted on glass slides, and specifically includes the staining of such biological specimens in specimens for microscopic examination. Staining and testing of biological specimens is performed in hospital laboratories, emergency rooms,
It occurs in large numbers every day in clinics, biology departments, blood banks and numerous physician's offices, veterinary clinics and hospitals. The slides used are made of glass and generally about 25
．． It has an area of 4mm x 76.2@ (1" x 5') and a thickness of 1mm. Some slides have a slightly different area of 25mm x 75mm or a different thickness of 1.2mm. To be stained and tested Samples include tissue, blood,
It may be saliva or urine. Biological specimens are commonly coated onto glass slides and air dried, often with heat or reagent fixation.

In order to stain a biological specimen fixed on a glass slide, it is necessary to bring a staining solution or reagent into contact with the specimen on the slide.

In one widely used prior art staining method, slides are placed horizontally, specimen side up, on a shelf suspended in a sink. Pour or drip stain from the bottle onto the slide to overflow the specimen. This technique wastes expensive staining fluid, since typically much more fluid is wrung out or dripped than is exactly needed for staining. Also, the sink becomes dirty and unsightly after some time from overflowing liquid. Workers' fingers also get dirty when picking up specimens.

If reagent bottles remain in the sink area for an extended period of time, the fluid will deteriorate or become contaminated.

Generally, each of the above techniques requires 6 to 8 rnl of reagent to perform an analyte test.

Another, less universal, known method is to place one or more slides vertically on a shelf and then to immerse the shelf with the slides into a container of staining solution. This method also uses much excess dye than is exactly needed for staining and often results in contamination when reagents are used on more than one slide. Similarly, this technique involves injection of liquid from a bottle into a container and injection of staining liquid from a container into a sink.

A sixth method, also less universal, is to use automated staining equipment. Large hospitals with heavy workloads tend to use this method. The mechanical equipment associated with such automated methods is extremely expensive and can only be justified if the dyeing quantities are very large. The cost of such machines precludes their use in many dyeing operations. Additionally, automated staining equipment is only available for one or two specimens.

The slide with fixed specimen is inserted vertically and longitudinally into a narrow chamber formed within the container. The slides are guided and positioned on the shelves in the room. When the slide displaces the liquid reservoir at the bottom of the chamber into the narrow space adjacent to the face of the slide, a thin film of staining solution forms on each side of the slide. The volume of staining solution used is approximately equal to the volume required to create a thin film on each side of the slide. This amount is as small as 1 ml.

When a stained slide is removed from the chamber vertically, the stain drips off the slide and returns to the sump at the bottom of the chamber. The container and working liquid can then be discarded.

The slide is inserted and removed from the chamber by a portion of the slide that extends continuously above the chamber.

In the present invention, 1y <
A small amount of staining solution is required; a new batch of reagent is prepared for each specimen;4 the staining solution stays in the chamber, so the sink doesn't get dirty; the operator's fingers don't get dirty, because the staining solution stays in the room; And since it is a prepack method, the method is quick and saves a lot of time.

The container is disposable and the lid or stopper seals the chamber and stain until use.

The present invention specifically utilizes a conventional slide 20 as shown in FIGS. 2-4. The slide 20 is simply a rectangular piece of glass 21.

A specimen 22 is placed above it for examination.

The slide has opposing flat surfaces 23 and 25, opposing long edges 26 and 27, and opposing short edges 28.

The most commonly used slides are approximately 7.62cIIL
(6 inches) long Ls), approximately 2.54 cIrL (1
It has a width (Ws) of about 1 inch and a thickness Ts of about 1 inch. In some cases the slide is 75mm Ls
and a Ws of 25 mm, or a Ts of 1.2 mm.

These dimensions vary slightly depending on the manufacturer. For example, the thickness is approximately 0.13 cIIL to 0.086 cr/L (
The width varies from 0.0525" to 0.32"), and the width varies from approximately 2°58 crrL to 2.45 boxes (1,016" to 9.61").
It can change to

Specimen 22 is prepared and applied to the surface of the slide in any known manner, such as by air drying, heat, or use of reagent fixation. The specimen 22 is generally positioned centrally on one of the slide surfaces, with a clean area free of specimen at each end of the surface.

Slide 20 with unstained specimen 22 is intended to be inserted into disposable slide staining device 30 . Apparatus 30 is preferably made of clear or translucent plastic and is generally rectangular in profile with a size that generally matches the size of slide 20. The device is intended for use in an upright position 9 and has a top surface 30, a bottom surface 32, a lip 33 and sides 35 and 36. A boxwood or flange 29 surrounds the top surface 30 and has a flat top surface.

The inner cavity or chamber 40 is likewise rectangular in profile and is also generally adapted to the dimensions of the slide 20. inner space 4
0 has opposing vertical tapered guide walls 43 and 45 along vertical edges 43 and 45 of the cavity extending from the top surface 46 to the bottom surface 47 of the cavity. The cavity also has opposing sides 48 and 50.

The cavity 40 has a beveled edge 52 at its upper end.

A lid or seal 55 is affixed to the upper surface 31 of the boxwood or hemlock 29 by any suitable means, such as a beat seal or an adhesive.

The lid is intended to be peeled off prior to use by pinching and pulling on the knotweed, so that the cavity 40 is exposed.

The staining liquid reservoir 0 is contained in the cavity, and extends to approximately the middle of the cavity, as shown in FIGS. 1 and 10.

The device according to the invention, as shown in FIG. 1, is intended to be extremely easy to manufacture with respect to the sealing of the staining solution.

As seen in FIGS. 5-8, the slide 20 is shown sequentially being inserted into the cavity 40. FIG. 5 shows a state in which a slide v40, to which a biological specimen 22 to be stained is attached, is entering a space between guide walls 41 and 42. The slide 20 has opposing sides 48, as best seen in FIGS. 15 and 16.
and 49.

The slide in FIG. 5 is just in contact with the upper surface of the staining solution reservoir 60, which is a static liquid level. The static liquid level is at a height H1 above the bottom of the cavity.

In Figures 6 and 7, the slide is progressively lowered into the cavity and into the stain reservoir, the fluid in the stain reservoir is displaced, and the dynamic fluid level is such that the slide is fully inserted. In this case, H2 is reached as shown in FIG. At this point, the stain completely covers the specimen 22. Hl is about 4 times H2. H2 also represents the height of the void, as best seen in FIG.

FIG. 9 diagrammatically illustrates the principle described and illustrated in FIGS. 5-8. If the slide plate sinks below the static surface, the dynamic level of the staining solution actually rises correspondingly in the linearization case.

If the slide is removed from the staining solution after it has been left for an appropriate amount of time to complete the staining, the slide will be pulled up and the reverse order will occur. The liquid level correspondingly falls from the side of the slide in the form of Hedren's liquid in the staining solution reservoir. There will be some residual stain adhering to the specimen and slide, and of course the height of the stain reservoir at the end of the staining will drop below the static liquid level as the slide is withdrawn from the instrument. . When the slide is withdrawn from the apparatus 30, it has a stained specimen 54, as shown in FIG. Since the device is intended for single use, any additional reduction in reservoir height and volume is not unpleasant.

A guide portion 51 with a tapered green or bevelled edge 52 and a beveled side 53 allows easy insertion of the slide into the guideway. This also serves as a vertical hole to collect overflow of staining solution as well as splashes from rapidly dropping slides into the cavity. A flap or guide portion 29 extends to the upper end or top shim of the device and has a flat top portion 63 which may be suitably secured thereto by suitable heat sealing techniques. The foil may be comprised of a metal or plastic material that can be peeled from the covering position by pinching the tab portion 56 and pulling it to the fully open position, as seen in FIG. The lid portion in the sealing and covering position is shown in FIGS. 10 and 11.

The width and thickness of the slide are variable, whatever the requirements or requirements to have special dimensions. By tapering the guideway, the device accommodates slides of variable thickness and variable width while still keeping the slides away from the interior walls of the cavity.

Additionally, the tapered guideway ensures that only the edges of the sly P contact the guideway, but not the surface bearing the analyte.

When the slide 20 is inserted into the cavity 40 by means of guideways 41 and 42, it assumes a very defined position and orientation within the cavity with respect to the sides or walls 48 and 49 of the cavity 4o.

As seen in Figures 14-16, slide 20
When inserted in its lowest position, the upper part 61 is connected to the device 3.
It extends above the cavity 40 and the top 31 of Ω. The upper portion 61 of slide 20 does not have analyte 22 since the analyte area is normally in the center of the slide, as indicated by the dashed line in FIG. The slide 200 portion 61 that remains above the device 30 is used to grip the slide for insertion into and removal from the device. The slide is also gripped by this part during the water wash following staining.

In this way, the operator's fingers do not come into contact with the staining solution and remain dye-free. Above the device 30, slide Ls
It extends by about % of its length.

Specimen 22 is stained especially when the slide is fully inserted as shown in FIGS. 14-16. As seen particularly in FIGS. 15 and 16, the slide is positioned in the center of the cavity and spaced from walls or sides 48 and 49. The slide 2o has gaps 65 and 66 adjacent to each of its faces containing a staining solution. The slide can be inserted with the specimen 22 facing either direction as the slide 20 enters the cavity. The surface of the slide 20 and the specimen 22 thereon prevent the specimen 22 from touching the sides of the cavity, but rather into the stain in the space 65 or 66 formed by the slide over the lance and the sides of the cavity. Positioned so as to remain exposed. A minimum amount of staining solution is needed to stain the specimen when the slide is positioned in this manner. This is clearly shown in the cross-sectional views of FIGS. 15 and 16.

From the above description, the space 6 adjacent to each side of the slide
5 and 66 are flat and thin gaps when the slide is fully inserted into the cavity, allowing an appropriate amount of staining to properly stain the specimen on the slide without using excessive amounts of staining solution. Contains liquid. Furthermore, the slide itself
It is used to position the stain adjacent to the face of the slide and the specimen on either side. It is clear that the direction in which the specimen faces when inserted into the cavity is not important since there is a liquid film on both sides of the slide.

The slide itself cooperates with the container to create a film of stain adjacent to the slide surface9, and the stain in these interstices is absorbed into the voids and reservoirs that form there when the slide is removed. It can flow to the bottom.

10-16 show the interior of the apparatus of FIG. 1 in detail. Ws is the width of the slide 20, 6D, Ls is its length, and Ts is its thickness. The height of the cavity as well as the height of the dynamic liquid level mentioned above, H2, is approximately % of the slide length Ls.

The guideways 41 and 42 have tapered walls at the edges of the cavity as shown. As can be seen in FIG. 16, the guideway width WT is also slightly larger than the slide width Ws, for example by about 0.043 cIIL (0.02 inches), so Appropriate clearance exists. The thickness TT at the tapered end of the guideway is slightly less than the slide thickness Ts, for example about 0.03 cm (0.01'6 inches) less. The thickness Y of the void is approximately 2 of the thickness Ts of the slide P.
It's double. By means of tapered guideways 41 and 42,
When inserted into the guideway, the slide lies centrally spaced between the side walls of cavities 48 and 49. If the thickness of the cavity is twice the thickness of the public slide, the thickness of the gap formed on each side of the slide when the slide is inserted is about % of the thickness of the slide P.

Due to the tapered guideways 41 and 42, slides of slightly varying dimensions, such as those described above, are suitably spaced within the cavity 40. 6. The tapered walls of guides 41 and 42 are used during positioning of the slide. Guiding with its edges compensates for these slight variations. Even when using a slide with a minimum width and/or thickness within variable dimensions, the slide will have a spacing from the sidewalls of the cavity, but all edges of the slide will not be in intimate contact with the guide. Although such relatively small slides have greater freedom of movement within the cavity than relatively larger slides, the restraining force imposed on the corners of the slide by the tapered walls of the guideway may cause the slide V to be spaced apart from the cavity. is kept in place so that a stain film can form adjacent to the slide surface.

It is clear that the above dimensions are merely exemplary to indicate a suitably sized container for practicing the invention.

In order to make the invention more fully clear, the present invention has been specifically described with reference to specific embodiments IC.
However, without departing from the spirit and scope of the invention as defined by the claims,
It will be clear that the invention may be implemented in various forms other than those specifically described and illustrated.

[Brief explanation of the drawing]

FIG. 1 is a perspective view of the slide staining device of the present invention, FIG. 2 is a perspective view of a conventional slide glass, and FIG. 3 is a perspective view of the device with the lid removed and a slide inserted. 4 is a similar perspective view, and FIG. 4 is a schematic representation of a conventional slide, such as that shown in FIG. 2, stained in the apparatus of the invention;
5, 6, 7, and 8 are vertical sectional views sequentially showing the slide inserted into the device of the present invention, and FIG. FIG. 10 is an enlarged sectional view taken along line 10-10 of FIG. 1, showing in detail the internal design of the dyeing device shown in FIG. 1, and FIG. 11 is a diagram showing the structure. 11-1 in Figure 10 showing the details of
12 is a cross-sectional view along line 1, and FIG. 12 shows details of the slide entrance hole on the top surface of the staining device.
12 is a plan view of the staining device directly below the foil lid, and FIG. 13 shows the outline and size proportions of the internal chamber of the staining device and the slide guide slot of FIG. 10;
3-13, and FIG. 14 is a cross-sectional view taken along the line 3-13, and FIG. 14 shows the slide completely inserted into the staining device as shown in FIG. FIG. 15 is an enlarged sectional view similar to FIG. 10, and FIG. 15 is an enlarged cross-sectional view of FIG. Figure 16 is a cross-sectional view taken along line 15, 16-16 of Figure 14 showing the relative size proportions and locations of the slide and slide staining volume
FIG. 20...Slide, 22...Specimen, 29...Brim, 30...Staining container, 31...Top surface of container, 32...
...Bottom surface, 35.36...Side surface, 40...Inner cavity, 41, 42...Guidance path, 54...Stained specimen, 55...Lid, 56...Tab, 60...Staining solution reservoir, 65.66...Gap

Claims (1)

[Claims] 1.1) moving the slide vertically downward relative to the container and to its bottom within the container; 2) displacing the liquid in the liquid reservoir upwardly relative to the container by the slide; a) the reservoir is substantially at the height of the container before the slide is moved vertically downward; A method for staining biological specimens on a microscope slide, characterized in that the slide is displaced upwardly over substantially the entire height of the container. 2.1) Contains a reservoir for the staining solution in the cavity, and the liquor reservoir has a moat at the height of the cavity; 2) The reservoir in the cavity can be simply displaced upward. 6) inserting the slide with the fixed specimen into the reservoir; and 4) displacing the reservoir upward along the slide substantially to the top of the cavity. Methods for staining biological specimens on microscope slides. 6.1) forming a reservoir of contained stain; 2) displacing the stain by vertically inserting a slide into the contained reservoir; and 3) displacing the displaced stain into the slide. A method for staining a biological specimen on a microscope slide, which is characterized by forming a liquid film that stains the specimen by restricting it along the surface of the slide. 4.1) move the slide downwards relative to the container containing the staining solution; and 2) displace the staining solution in the solution reservoir upwardly relative to the container, with the liquid level facing upwards. A method for staining a biological specimen on a microscope slide, characterized in that the specimen is moved approximately twice the height of the slide. 5. To form a liquid film of staining solution on the surface of the slide,
The method of claim 1, wherein the slide is moved downwardly into a reservoir of staining solution, thereby displacing the reservoir upwardly and forming a film of fluid on each side of the slide. , an apparatus for staining with a staining solution a biological specimen fixed to one of the opposing sides of a microscope slide, comprising: a) a cavity; and b) a housing for receiving the slide when the slide is inserted into the cavity; a guideway extending vertically within the cavity for guiding; C) a gap within the cavity adjacent each side of the slide when the slide is inserted into the cavity; and d) a volume of staining solution within the cavity. and a container with a staining solution forming a reservoir at the bottom of the cavity before inserting the slide;
A device for staining biological specimens on a microscope slide, characterized in that the staining solution is displaced into a gap adjacent to the surface of the slide when the slide is inserted into the cavity by a guideway. The Z container has a reservoir volume approximately equal to the gap adjacent to the surface of the slide.1. A dyeing device according to claim 6. 8. The staining device according to claim 6, wherein the container is such that when the slide is inserted completely into the cavity, the gap adjacent to the surface of the slide with the fixed specimen covers the specimen. 9. The staining device according to claim 6, wherein the container has a flat gap adjacent to the surface of the slide and a thickness smaller than the thickness of the slide. 10. The staining apparatus according to claim 9, wherein the container has a gap approximately equal in thickness to the thickness of the slide. and the container is arranged such that the liquid reservoir at the bottom of the cavity is displaced to a height approximately twice the height of the liquid reservoir before inserting the slide upwardly into the gap. Staining equipment as described in Section. 12. The staining apparatus according to claim 6, wherein the container has a height smaller than the length of the slide. 13. The container has a height smaller than the length of the slide. 13. The container has a height smaller than the length of the slide. 7. A dyeing device according to claim 6, which extends beyond the dyeing device. (4) 14. The dyeing device according to claim 6, wherein the guide path has a tapered cross section. 15. 1) a container; 2) a guideway fixed within the container and capable of vertically moving the slide to a downward position;
) surrounding the guideway respectively forming a) a volume confined around the guideway and b) a gap on each side of the slide within the confined volume when the slide is in a downward position in the guideway. 4) a cavity in the container having walls; and 4) a reservoir for staining solution located at the bottom of the guideway and displaced into the gap on each side of the slide by the slide in its downward position. A staining device for biological specimens on microscope slides, characterized by: 16. The apparatus of claim 15, wherein the gap on the surface of the slide is flat and covers the specimen on the slide. 1Z The device according to claim 15, wherein the guide path has a tapered cross section. 18. The device of claim 16, wherein the thickness of the gap adjacent the face of the slide is less than the thickness of the slide. 19. The device of claim 18, wherein the thickness of the gap adjacent the face of the slide is approximately equal to a spoonful of the thickness of the slide. 20. 1) a container; and 2) a container adapted to accommodate a slide within the container, having a) a width approximately equal to the width of the slide, and b) a thickness approximately equal to twice the thickness of the slide. 3) a slide within the container having a slide in the cavity, a) the width of the slide extends to the width of the cavity, and b) the thickness of the slide is centered in the thickness of the cavity. 1. A staining device for biological specimens on a microscope slide, comprising a guide path for guiding and positioning the specimen. 21. The device according to claim 20, wherein the guide path has a tapered cross section. 22. The height of the void is approximately equal to 4 of the length of the slide,
An apparatus according to claim 20. 26. The device of claim 20, wherein the slide features a flat gap adjacent each side of the slide when inserted into the cavity. 24. The device of claim 20, wherein the planar gap has a thickness approximately equal to the thickness of the slide. 25. The device of claim 20 having a removable seal covering the cavity. 26. The device according to claim 6, which has a staining solution reservoir in the cavity. 2Z The device of claim 26, having a removable seal covering the cavity. 28. 1) a container; 2) a first gap in the container for accommodating a slide and parallel to and adjacent to the first gap for containing a flat liquid film when the slide is in the container; 3) a guide path for guiding the sliver r into the first gap within the container, the second gap having a thickness smaller than the first gap; and a second gap covers the specimen on the surface of the slide when the slide P is in the container (7). 29. The device of claim 28, wherein the guideway has a tapered cross section. 60, having a volume of staining liquid forming a puddle at the bottom of the cavity when the slide is outside the cavity and extending into the second gap when the slide is inside the cavity; Device. 31. 1) a) a cavity with a rectangular cross-section, b) a container with a height smaller than the height of the slide, and 2) a container within which the slide is guided vertically into the cavity and from which it escapes. 6) a liquid reservoir at the bottom of the cavity, the liquid reservoir surface being substantially % of the height of the cavity when the slide is outside the cavity; An apparatus for staining biological specimens on a microscope slide, characterized in that when the slide is completely inserted, the liquid reservoir is displaced to the top of the cavity.