CA1200744A - Method and structure for staining of biologic slides - Google Patents
Method and structure for staining of biologic slidesInfo
- Publication number
- CA1200744A CA1200744A CA000426723A CA426723A CA1200744A CA 1200744 A CA1200744 A CA 1200744A CA 000426723 A CA000426723 A CA 000426723A CA 426723 A CA426723 A CA 426723A CA 1200744 A CA1200744 A CA 1200744A
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- slides
- slide
- spacers
- cavity
- staining
- Prior art date
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Abstract
Abstract of the Disclosure Method and apparatus for neatly and efficiently coating microscope slides with a staining liquid. One or more slides are dipped into a pool of liquid partially fill-ing a chamber in a container. The chamber has a cross-section conforming closely to the slide exterior so that slide im-mersion causes the liquid level to rise upwardly and cover all those portions of the slides which may contain a specimen to be stained. To stain a multiplicity of slides simul-taneously, the slides are clamped in a holder between spacers which hold them parallel and spaced apart by a small predetermined distance, and the clamped slides are then held by the holder during subsequent immersion and removal. A
special inexpensive arrangement of case and slidable drawer with wedge-shaped side portions is used to facilitate fast clamping and unclamping of the slides.
special inexpensive arrangement of case and slidable drawer with wedge-shaped side portions is used to facilitate fast clamping and unclamping of the slides.
Description
METHOD AND STRUCTURE FOR
STAINING OF BI_LOCIC SLIDES
Background of the Invention Field of the Invention The invention generally relates to microscopic exarnination of biologic specimens mounted on glass slides, and specifically involves the staining of such biologic specimens in preparation for microscopic examination. The staining and examining of biologic specimens occurs daily in great numbers in hospital laboratories, emergency rooms, clinics, biology departments, blood banks, and many physi-cians' offices, ve-terinarian offices and hospitals. The slides used are of glass and generally have a 1" x 3" face dimension, and a lmm thickness. Some slides have a slightly different face dimension of 25mm x 75mm or a different thick-ness of l.2mm. The specimens to be stained and examined may, for instance, be tissue, blood sputum, or urine. The biologic specirnents are usually smeared on the glass slide and leEt to air dry, often with heat or reagent fixation.
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STAINING OF BI_LOCIC SLIDES
Background of the Invention Field of the Invention The invention generally relates to microscopic exarnination of biologic specimens mounted on glass slides, and specifically involves the staining of such biologic specimens in preparation for microscopic examination. The staining and examining of biologic specimens occurs daily in great numbers in hospital laboratories, emergency rooms, clinics, biology departments, blood banks, and many physi-cians' offices, ve-terinarian offices and hospitals. The slides used are of glass and generally have a 1" x 3" face dimension, and a lmm thickness. Some slides have a slightly different face dimension of 25mm x 75mm or a different thick-ness of l.2mm. The specimens to be stained and examined may, for instance, be tissue, blood sputum, or urine. The biologic specirnents are usually smeared on the glass slide and leEt to air dry, often with heat or reagent fixation.
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- 2 The Prior Art To stain a biologic specimen fixed on a glass slide, it is necessary to bring a staining liquid or reagen-t into contact with the specimen on the slide. For the presen~ purposes it will be understood that the term stain-ing includes application -to the specimen of any of various liquids or reagents during a staining process, even if a par-ticular liquid does not itselE produce the actual staining.
In one widely used prior art method of staining, the slide is placed horizontally, specimen up, on a rack sus-pended in a sink. The staining liquid is poured or dripped from a bottle over the slide to flood the specimen. This technique wastes expensive staining liquid, since much more liquid is generally squeezed or dripped than is actually needed for stainingO Also, the sink becomes stained and un-sightly after a period oE time from the overflow liquid.
The operator's fingers become stained when the specimen is picked up. When the bot-tle reagents remain in the sink area for a prolonged period of time, the fluid can deteriorate or become contaminated, intentionally or accidentallyO
It generally re~uires from 6 cc to 8 cc of each reagent to perform a specific examination with the above technique.
A less common alternative prior art method is to place the slide or slides vertically in a rack, and then to dip the rack with the slide into a container of staining liquid. This too uses an amount of dye far in excess of that actually necessary for staining, and, where the reagent is used for more than one slide, contamination often occurs.
Likewise, this technique involves pouring liquid from bottles into containers and pouring the staining liquid from containers into sinks.
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In one widely used prior art method of staining, the slide is placed horizontally, specimen up, on a rack sus-pended in a sink. The staining liquid is poured or dripped from a bottle over the slide to flood the specimen. This technique wastes expensive staining liquid, since much more liquid is generally squeezed or dripped than is actually needed for stainingO Also, the sink becomes stained and un-sightly after a period oE time from the overflow liquid.
The operator's fingers become stained when the specimen is picked up. When the bot-tle reagents remain in the sink area for a prolonged period of time, the fluid can deteriorate or become contaminated, intentionally or accidentallyO
It generally re~uires from 6 cc to 8 cc of each reagent to perform a specific examination with the above technique.
A less common alternative prior art method is to place the slide or slides vertically in a rack, and then to dip the rack with the slide into a container of staining liquid. This too uses an amount of dye far in excess of that actually necessary for staining, and, where the reagent is used for more than one slide, contamination often occurs.
Likewise, this technique involves pouring liquid from bottles into containers and pouring the staining liquid from containers into sinks.
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3 A third, again less common procedure is to use
4 an automated staining device. Large hospitals with heavy workloads are more inclined to use this method. The 6 machinery involved in such automated method is extremely 7 expensive and can only be justified where the amount of B staining is extremely heavy. The cost of such machines 9 prohibits their use in most staining operations. Further-more, automated staining devices are only available for 11 one or two stains.
13 Summary of the Present_Invention 14 In our co-pending Canadian application serial number 426,724, April 26, 1983 for METHOD AND STRUCTURE
16 FOR ST~INING OF BIOLOGIC SLIDES, a slide with a specimen 17 fixed thereon is inserted vertically, lengthwise, into a 18 confined chamber formed within a container. The slide is 19 preferably guided and positioned within the chamber on tracksO A thin film of staining liquid is formed on each 21 face of the slide when the slide displaces a pool at the 22 bottom of the chamber into confined spaces adjacent to 23 the faces of the slide. The volume of staining liquid 24 used is approximately equivalent to the amou~t of liquid necessary to form the thin film on each face of the 26 slide. This amount can be as little as 1 cc.
28 ~hen sealed, the container is rendered tamper-29 proof so that the contents cannot be adulterated prior to use, thus preventing a false reading of the stained 31 slide.
33 When the stained slide is vertically withdrawn 34 from the chamber, the staining liquid drains off the slide and retuxns to a pool at the bottom of the chamber.
36 The container and used liquid can then be disposed of.
37 The slide may be inserted and withdrawn from the chamber, 38 and subse~uently handled, by a portion of the slide which 39 continuously extends above the chamber.
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Substantially less stainin~ liquid is required than in the prior art; a fresh batch of reagent is provided for each specimen; sinks are not stained since the liquid remains within the chamber; the operators' fingers do not become stained, since the staining liquid remains within the chamber; and, since the stain is prepackaged, the method is quicker and more time eficient.
The container is a disposable one intended for one use, and has a cover or stopper which seals the chamber and liquid until there is such use. The container is render-ed tamperproof giving assurance that the staining process will yield a true result. After the staining is complete, the chamber may be easily resealed before disposal to avoid any subsequent spattering or leakage of stain f om the cham-ber.
The above embodiment relates to a single slide,while the embodiment now to be described relates to simul-taneously staining a m~lltiplicity of slides.
The inventive concept of utilizing a thin film of staining liquid on each face of the slide, wherein the film is formed from a limited pool of staining liquid which is displaced by the immersion of the slide itself into the container, remains the same as in the prior application referred to above, but the present embodiment permits the basic concept to be used with a multiplicity of slides.
The present invention uses a holder that posi-tions, spaces and secures slides with respect to one another.
The spacing and positioning of the slides in the holder is critical to achieving the effect sought in the invention, namely a thin film on the face of each slide which is formed by the slides themselves in cooperation with the container when the slides are immersed into a limited pool which occu-pies only a portion of the container when the slides are not immersed therein. The slides, while spaced, positioned and secured in the holder, are inserted into a special, (`7~
matching, container holding a limited pool of staining liquid.
~he slides displace li~uid of the pool, in the fashion set forth in the parent application, so that a thin fil~ is formed on the faces of the slides.
When the slides, by means of the holder, are with-drawn from the containe~, the ilms of staining fluid fall back into the container and reform substantially the same pool as originally formed.
The slides are then removed from th~ holder for further processing.
The holder, container, and pool are specifically designed for a designated number of slides, and it is intended that, in the preferred use of the system, no more and no less than the designated number of slides be used in and with that particular container, holder, and pool.
As with the single slide embodiment disclosed in the prior application, preferably the container is ini-tially sealed with the staining fluid therein, and is re-sealed after use and before discard. The holder is, de-sirably, removably attached to the container prior to use,so that the elements of the invention are presented together for use in combination.
Of course, a cooperating container, holder, and staining liquid pool can be prod~ced for any plurality of slides within reason, but it is contemplated that the most popular use w~ll be a five-slide embodiment.
Brief Description of the Drawin~s Fig. 1 is a perspective view of a slide staining device.
Fig. 2 is a pers~ective view of a conventional glass slide of either metric or ~nglish dimensions.
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Fig. 3 is a ~erspective view similar to Fig. 1 showing the slide staining device with the cover removed and a slide inserted.
Fig. 4 i~ a view of a conventional slide such as shown in Fig. 2, haYing been stained in the device of Fig. 1.
Fiqs. 5 through 8 are sectional elevational views showing a slide in successive stages of its insertion into the device of Fig. 1 of the invention.
Fig. 9 is a graph illustrating a principle involved in the steps of Figs. 5 through 8.
Fig. 10 is an enlarged sectional elevational view taken on the line 10 10 of Fig. 1~ showing in greater detail the interior design of the staining device of Fig. 1.
Fig. 11 is a ~ransverse sectional view taken on the line 11-11 of Fig. 10, showing still further details of construction.
Fig. 12 is a plan view of the staining device directly beneath the foil cover taken on the line 12-12 of Fig. 10~ showing details of the slide entrance aperture on the upper face of the staining device.
Fig. 13 is a sectional plan view taken on the line 13-13 of Fig. 10, showing the profiie and proportions of the i~terior chamber and slide~guiding slots of the stain-ing device.
Fig. 14 is an enlarged sectional elevational viewsimilar to Fig. 10, but showing a slide fully inserted in the staining device, such as is shown in Fig. 3, and showing the combined propo~tions and the positions of the slide and slide-staining device in use.
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Fiy. 15 is a transverse sectional elevational view taken on the line 15-15 of Eig. 14 showing central positioning of a slide carried in slots and showing liquid displacement chambers formed Oll the front and rear faces of the slide.
Fig. 16 is a sectional plan view taken on the line 16-16 of Fig. 14, showing relative proportions and positions of the slide and slide-staining device, and liquid displacement chambers formed on the front and rear sides of the slide.
Fig. 17 is a perspective view of the multiple slide staining tank of this invention, with the seal cover partially removed.
Fig. 18 is an exploded perspective view showing both the slide holder and staining tank of the invention.
Fig. 19 is a perspective view of both the slide holder and staining tank in a co-acting condition in accord-ance with this invention.
Fig. 20 is a perspective view of the empty slide holder of this invention prior to loading.
Fig. 21 is an exploded perspective view of the slide holder shown in Fig. 2Q showing all the co-acting elements of the slide holder of this invention.
Fig. 22 is a perspective view of the slide holder of Figs. 20 and 21 being loaded with five slides to be stained.
Fig. 23 is a sectional elevational view taken on the line 23-23 of Fig. 22, showing the five slides loaded in the holder, prior to clamping thereirl.
Fig~ 24 is A sectional plan view taken on the line 24-24 of Fig. 23 showing additional details of the five loaded slides loosely held by the slide holder.
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Fig. 25 is a sectional elevational view similar to Fig. 23 but showing the five slides clamped in tightly held spaced relation~hips, and with the five slides extend-ing in planes normal to the top and bottom faces of the slide holder.
Fig. 26 is a view simil~r to Fig. 24, ~ut showin~
the slides in a clamped position.
Fig. 27 is a side sectional elevational view of the slide staining tank taken on the line 27~27 of Fig.
17, showing interior details of the tank.
Fig. 28 is a front sectional elevational view of the slide staining tank taken on the line 28-28 of Fig.
17, showing additional details of the tank interior.
Fig. 29 is a side sectional elevational view taken on the line 2~-29 of Fig. 19, showing details of the tank interior when in engagement with the loaded slide holder.
FigO 30 is a front sectional elevational view taken on the line 30-30 of Fig. 29, showing additional details of the tank interior when engaged with the loaded slide holder and showing the resultant upward displacement of a staining fluid about the slides.
Des~ri~tion of the Preferred Embodiments A conventional slide 20 is shown particularly in Figs. 2 and 4. The slide 20 is simply a rectangular piece of glass 21 on which a specimen 22 is mounted ~or microscopic examination. The slide has opposed flat sur-aces or faces 23 and 25, opposed long edges 26 and 27, and opposed short edges 28.
The most commonly used slide has a length (Ls) of approximately 3", a width (Ws) of approximately 1", and a thickness (Ts) of approximately lmm. In some instances, the slide may have a LS of 75mm and Ws of 25mm, or a TS
of 1.2mm.
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. 9 qlhese dimensions may vary slightly with different manufacturers. For instance, thicknesses may vary from .0525" to .0325~, and the width may vary from l.Q16" t~
.964".
The specimen 22 is prepared and then affixed to a face of the slide in any well known manner such as by use of air drying, heat, or reagent ~ixation. The specimen 22 is generally positioned at the center of one of the slide faces, with a clear zone, free of the specimen, on each end of the face. The specimen is virtually invisible at this point, prior to staining.
The slide 20, with the unstained specimen 22 thereon, is intended to be inserted into a disposable slide-staining device 30~ Device 30 is desirably formed of a transparent or translucent plastic into a generally long rectangular configuration having proportions yenerally conforming to the proportion of ~he slide 20. The device is intended to be used vertically, and has a top 31, a bottom 32, edges 33 and sides 35 and 36 A collar or flange 29 surrounds top 30 and has a flat surface thereon.
An internal cavity or chamber 40, likewise is rectangular in configuration and again, conforms generally to the proportions of the slide 20. The internal cavity 40 has opposed tapered vertical tracks 41 and 42, along the vertical edges 43 and 45 of the cavity extending from the top 40 of the cavity to the bottom 47. The cavity also has opposed sides 48 and 50.
The cavity 40 has at its upper end a sloping~
downwardly converging portion 51 having beveled edges 52 and sides 53.
-1,0-A cover or seal 55 is a~fixed to the top 31 on collar or flange 29 by suitable means, such as heat sealing or adhesive. The cover is intended to be peeled back by grasping and pulling on tab 56 prior to use, so that the cavity 40 is exposed A pool of staining liquid 60 is contained within the caYity and extends about halfwa~ up the cavity, as seen in Figs. 1 and 10.
The device is intended to be manufactured at a central facility whereat it is sealed with the staining liquid therein, as seen in ~ig. 1.
As seen in Figs. 5 through 8, there is shown in sequence views of a slide 20 being inserted into the cavity 40. In FigO 5, the slide 40 with a biological specimen 22 to be s~ained af~ixed thereon, is shown entering the cavity 40 on tracks 41 and 42. The slide 20 i5 spaced from the opposing sides 48 and 49 as seen best in Figs. lS and 16. The slide in Fig. 5 is just about to contact the upper surface of the pool 60, which is the static liquid level.
20 The static liquid level is at a heighth of Hl above the bottom of the cavity.
In Figs. 6 and 7, the slide progressively descends into the cavity below the pool, the liquid in the pool is displaced and rises until the dynamic fluid level reaches H2 as seen in Fig. 8, when the slide is fully inserted.
At this point, the staining liquid entirely covers specimen 22. Hl is approximately one~half of ~2. H2 also represents the heighth of the cavity as seen best in Fig. 14.
In Fig. 9~ the principle described and shown in Figs. 5 through 8 is illustrated in yraph form. As the slide is submerged below the static liquid level, there is a corresponding rise in the dynamic liquld level of the staining liquid in virtually a straight line relationship.
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As the slide is removed from the staining li~uid, after being left therein for an adequate period to accomplish staining, the slide is withdrawn and the reve~se s~quence occurs. The liquid level alls correspondingly as the liquid drains from the side of the slide into the pool. There is some residual staining liquid that adheres to the speci-men and the slide, and of c~urse the pool heighth at the end of staining drops correspondingly below the static liquid level, when the slide is withdrawn from the device. The slide when withdrawn from the device 30 has thereon the stained specimen 54 as seen in Fig. 4. Since the device is intended for a one-time use, such drop in the po~l heighth and volume is not objectionable.
The tapPred or guide portion 51 with beveled edges lS 52 and beveled side 53 permits easy insertion of the slide into the tracks. It also acts as a well to catch any over-flow of the staining liquid, including any splashing where the slide is dropped rapidly into the cavity. A collar or guide portion 29 extends around the upper end or top f the device and is integral therewith. Flange 29 has desirably a flat top portion 63 which permits a foil or cover 55 to be suitably secured thereto as by adhesive, or by a s~itable heat-sealing technique. The foil can be of a metallic or a plastic material capable of being peeled back from the covering by grasping a tab portion 56 and pulling to a completely open position, as seen in Fig. 3.
The cap portion in sealing and covering position is shown in FigsO 10 and 11.
There is variability in the width and thickness f slidesl even those claiming to have specific dimensions.
By tapering the tracks, the device will receive a slide of variable thickness and variable width and still keep the slide away from the inside wall of the cavity. Addi-tionally, the tapered tracks assure that only the edge of the slide will touch the track and not the surface which contains the specimen.
The slide 20 when inserted into the cavity 40 on ~he tracks 41 and 42 assumes a very definite position and posture within the cavity and witi- resp~ct to the sides or walls 48 and 49 of the cavity 40.
As seen in Fi~s. 14 to 16 inclusive, the slide 20 when inserted into its ~ottommost position has a top portion 61 which extends above the cavity 40 and top 31 of the device 30. The top portion ~1 of the slide 2D is clear of any specimen 22, since the specimen zone is conven-tionally in the middle of the slide as shown in Fig. 14 hy phantom lines. The portion 61 of the slide 20 remaining above the device 30 is used to grasp the slide for inserting into the device and for withdrawing the slide from the device.
The slide is also grasped by this portion during the rinsing subsequent to the staining. In this way, the operator's fingers remain free of contact with the staining solution, and free of the dye. Approximately one-sixth of the length of ~he slide LS extends above the device 30.
The spec;men 22 is stained when the slide is fully inserted as shown particularly in Figs. 14 through 16 inclu-sive. The slide as seen particularly in Figs. 15 and 16 i5 positioned centrally of the cavity 40 and spaced from walls or sides 48 and 49. The slide 20 has adjacent to each face thereof spaces 65 and 66 which contain staining liquid. The slide can be inserted with the specimen 22 facing in either direction as the slide 20 enters the cavity.
The face of the slide 20 and the specimen 22 thereon is positioned so that the specimen 22 cannot contact the sides o~ the cavity, but rather the specimen is held in a way that exposes it to the staining liquid in the space 65 or 66 formed ~y the slide and the side of the cavity. A minimum amount of stainin~ llquid is necessary to stain the specimen when the slide is so positioned. This is clearly shown in the sectional views in Figs. 15 and 16.
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~13-It will be seen from the above description that the space 65 anc7 66 adjacent to each face of the slide when the slide is fully inserted into the cavity is a flat thin space which contains a suitable amount of staining liq~id to properly stain the specimen thereon, without excess use of such liquid. ~rthermore, the slide itself is used to position this liq~id adjacent to the faces of the slide, and the specimen on one of the faces. It will be seen that it does not matter as to which direction the specimen ~aces when inserted into the cavity, since there is film on both faces of the slide.
The slide itself, in cooperation with the container, creates the film adjacent to the slide faces for staining, and then permits the liquid in those spaces to drain back lS into the bottom o~ the cavity and pool formed therein when the slide is removed.
In Figs. 10 throu~h 16 inclusive, there is shown in detail the interior of the device of Fig. 1. Ws is the width of the slide ~l LS is its length, and TS is its thick-ness~ ~2~ which is the heighth oE ~he cavity, as well asthe heighth of the dynamic liguid level discussed above, is approximately three~quarters of the slide len~th Ls~
The teacks 41 and 43 comprise tapered walls in the edges of the cavity, as shown. The width between the 2~ tracks, WT, as seen in Fig. 13! is sli~htly greater, for instance, .02~ greater, than the width of the slide Ws, so that there is adequate clearance for the slide to move in the tracks. TT, the thickness of the track at its tapered end, is slightly less, for instance, .016'i less, than the 30 thickness of the slide Ts~ Y, the thickness oF the cavity, is approximately twice the slide thickness, Ts. By virtue of the tapered tracks 41 and 42, the slide, when inserted în the tracks, is centr~lly spaced between the side walls ,~f;~ 7~
of the cavity, 48 and 49. Where the thickness of the cavity is twice that of the sli.de, the thickness of the spaces formed on each face of the slide when the slide is inserted will be approximately one-half the thickness o the slide.
By virtue of the tapered tracks 41 and 42, slides with slightly varying dimensions ~s set forth above will be suitably spaced within cavity 40. The tapered walls of the tracks 41 and 42 will compe~sate for these slight variations by guiding the slide at its edges thereof while still positioning the slide. Even with a slide having the smallest width andJor thickness within the variable dimen-sions, the slide is still spaced from the sides of the cavity, although all edges of the slide may not be in firm contact with the tracks. Even though such a relatively small slide ~ay have more freedom to move about within the cavity than a relatively large slide, the restraint imposed on the corners of the slide by the tapered walls of the tracks keeps the slide spaced from the cavity walls so that-a film of staining fluid can be formed adjacent the slide faces.
It should be understood that the dimensions given above are merely illustrative to indicate a suitable dimen-sioned container to practice the invention~
~eferring now to the preferred embodiment of the invention ~or simultaneously staining a multiplicity of slides, and with partic~lar reference to Figs. 17-30, slides 120 are of t.he nature, size and design described above with respect to slide 20.
The containe 130 may be genexally similar to the container 30 described above, in shape, heighth, and 30 width, except the horizontal depth of the container is ex- ~ :
tended to receive a plurality of slides, rather than onl~
one, as will be described. The container 130 has an open top 131, a bottom 132, edges 133, 134, sides 135, and 136, and collar or flange 129.
The internal cavity or main chamber 140 of con-tainer 130, has fo~r tracks s~ch as 141 and 142 in its four vertical corners, vertical edges 143 and 145, a bottom 1~7, and opposed sides 14B and 150.
In the version shown, the main chamber has at its top an overflow chamber 160 formed in the flange portion 129 which has side walls 168,170 formed by sides of the mai~ chamber 140, end walls 172,174 and bottom ledges such as 180.
~0 A seal cover 186, similar to seal 55 described above, is affixed to top 131 of the container.
The slides are spaced, positioned, and securely held in holder 200, which consists essentially of a drawer 202, a case 204, and spacers such as 206. The spacers are slidably and pivotally suspended in case 204 on pivot trun-nion pins such as 210 which ride in tracks 212 and 214 on the interiors of the opposed sidewalls 215,216 of the case.
Drawer 2~2 slides into case 204 so that when moved inwardly it pushes aqainst at least the foremost spacer 217~
The case 204 has a closed top wall 2~8 and an open bottom. (The case is shown inver~ed, in the loading position, in Figs. 20, 21, 22, 23 and 25. ) The side walls 215~216 contain respective slots 224 and 226, an open front and a closed back wall 230.
The tracks 212 and 214 in the case 204 are each formed by parallel ridges which deine channels 232, 234 for receiving the pins 210 on the spacers.
The spacers are thin, flat rectangles which pre-ferably have, at their long edges nearest the pins, a tapered edge or point for facilitating insertion between them of one end of the slides~ The pins are near the edges o~ the spacers so the spacers hang free and loose in the trAcks before loading of the slides. Preferably, these spacers are all alike.
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The drawer 202 has a front wall 300, push and pull knobs or ears 302, 304 at the opposite sides of the front wall, sides 306,308, and diagonal interior braces 310, 312 extending b~tween the front wall and the side walls
13 Summary of the Present_Invention 14 In our co-pending Canadian application serial number 426,724, April 26, 1983 for METHOD AND STRUCTURE
16 FOR ST~INING OF BIOLOGIC SLIDES, a slide with a specimen 17 fixed thereon is inserted vertically, lengthwise, into a 18 confined chamber formed within a container. The slide is 19 preferably guided and positioned within the chamber on tracksO A thin film of staining liquid is formed on each 21 face of the slide when the slide displaces a pool at the 22 bottom of the chamber into confined spaces adjacent to 23 the faces of the slide. The volume of staining liquid 24 used is approximately equivalent to the amou~t of liquid necessary to form the thin film on each face of the 26 slide. This amount can be as little as 1 cc.
28 ~hen sealed, the container is rendered tamper-29 proof so that the contents cannot be adulterated prior to use, thus preventing a false reading of the stained 31 slide.
33 When the stained slide is vertically withdrawn 34 from the chamber, the staining liquid drains off the slide and retuxns to a pool at the bottom of the chamber.
36 The container and used liquid can then be disposed of.
37 The slide may be inserted and withdrawn from the chamber, 38 and subse~uently handled, by a portion of the slide which 39 continuously extends above the chamber.
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Substantially less stainin~ liquid is required than in the prior art; a fresh batch of reagent is provided for each specimen; sinks are not stained since the liquid remains within the chamber; the operators' fingers do not become stained, since the staining liquid remains within the chamber; and, since the stain is prepackaged, the method is quicker and more time eficient.
The container is a disposable one intended for one use, and has a cover or stopper which seals the chamber and liquid until there is such use. The container is render-ed tamperproof giving assurance that the staining process will yield a true result. After the staining is complete, the chamber may be easily resealed before disposal to avoid any subsequent spattering or leakage of stain f om the cham-ber.
The above embodiment relates to a single slide,while the embodiment now to be described relates to simul-taneously staining a m~lltiplicity of slides.
The inventive concept of utilizing a thin film of staining liquid on each face of the slide, wherein the film is formed from a limited pool of staining liquid which is displaced by the immersion of the slide itself into the container, remains the same as in the prior application referred to above, but the present embodiment permits the basic concept to be used with a multiplicity of slides.
The present invention uses a holder that posi-tions, spaces and secures slides with respect to one another.
The spacing and positioning of the slides in the holder is critical to achieving the effect sought in the invention, namely a thin film on the face of each slide which is formed by the slides themselves in cooperation with the container when the slides are immersed into a limited pool which occu-pies only a portion of the container when the slides are not immersed therein. The slides, while spaced, positioned and secured in the holder, are inserted into a special, (`7~
matching, container holding a limited pool of staining liquid.
~he slides displace li~uid of the pool, in the fashion set forth in the parent application, so that a thin fil~ is formed on the faces of the slides.
When the slides, by means of the holder, are with-drawn from the containe~, the ilms of staining fluid fall back into the container and reform substantially the same pool as originally formed.
The slides are then removed from th~ holder for further processing.
The holder, container, and pool are specifically designed for a designated number of slides, and it is intended that, in the preferred use of the system, no more and no less than the designated number of slides be used in and with that particular container, holder, and pool.
As with the single slide embodiment disclosed in the prior application, preferably the container is ini-tially sealed with the staining fluid therein, and is re-sealed after use and before discard. The holder is, de-sirably, removably attached to the container prior to use,so that the elements of the invention are presented together for use in combination.
Of course, a cooperating container, holder, and staining liquid pool can be prod~ced for any plurality of slides within reason, but it is contemplated that the most popular use w~ll be a five-slide embodiment.
Brief Description of the Drawin~s Fig. 1 is a perspective view of a slide staining device.
Fig. 2 is a pers~ective view of a conventional glass slide of either metric or ~nglish dimensions.
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Fig. 3 is a ~erspective view similar to Fig. 1 showing the slide staining device with the cover removed and a slide inserted.
Fig. 4 i~ a view of a conventional slide such as shown in Fig. 2, haYing been stained in the device of Fig. 1.
Fiqs. 5 through 8 are sectional elevational views showing a slide in successive stages of its insertion into the device of Fig. 1 of the invention.
Fig. 9 is a graph illustrating a principle involved in the steps of Figs. 5 through 8.
Fig. 10 is an enlarged sectional elevational view taken on the line 10 10 of Fig. 1~ showing in greater detail the interior design of the staining device of Fig. 1.
Fig. 11 is a ~ransverse sectional view taken on the line 11-11 of Fig. 10, showing still further details of construction.
Fig. 12 is a plan view of the staining device directly beneath the foil cover taken on the line 12-12 of Fig. 10~ showing details of the slide entrance aperture on the upper face of the staining device.
Fig. 13 is a sectional plan view taken on the line 13-13 of Fig. 10, showing the profiie and proportions of the i~terior chamber and slide~guiding slots of the stain-ing device.
Fig. 14 is an enlarged sectional elevational viewsimilar to Fig. 10, but showing a slide fully inserted in the staining device, such as is shown in Fig. 3, and showing the combined propo~tions and the positions of the slide and slide-staining device in use.
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Fiy. 15 is a transverse sectional elevational view taken on the line 15-15 of Eig. 14 showing central positioning of a slide carried in slots and showing liquid displacement chambers formed Oll the front and rear faces of the slide.
Fig. 16 is a sectional plan view taken on the line 16-16 of Fig. 14, showing relative proportions and positions of the slide and slide-staining device, and liquid displacement chambers formed on the front and rear sides of the slide.
Fig. 17 is a perspective view of the multiple slide staining tank of this invention, with the seal cover partially removed.
Fig. 18 is an exploded perspective view showing both the slide holder and staining tank of the invention.
Fig. 19 is a perspective view of both the slide holder and staining tank in a co-acting condition in accord-ance with this invention.
Fig. 20 is a perspective view of the empty slide holder of this invention prior to loading.
Fig. 21 is an exploded perspective view of the slide holder shown in Fig. 2Q showing all the co-acting elements of the slide holder of this invention.
Fig. 22 is a perspective view of the slide holder of Figs. 20 and 21 being loaded with five slides to be stained.
Fig. 23 is a sectional elevational view taken on the line 23-23 of Fig. 22, showing the five slides loaded in the holder, prior to clamping thereirl.
Fig~ 24 is A sectional plan view taken on the line 24-24 of Fig. 23 showing additional details of the five loaded slides loosely held by the slide holder.
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Fig. 25 is a sectional elevational view similar to Fig. 23 but showing the five slides clamped in tightly held spaced relation~hips, and with the five slides extend-ing in planes normal to the top and bottom faces of the slide holder.
Fig. 26 is a view simil~r to Fig. 24, ~ut showin~
the slides in a clamped position.
Fig. 27 is a side sectional elevational view of the slide staining tank taken on the line 27~27 of Fig.
17, showing interior details of the tank.
Fig. 28 is a front sectional elevational view of the slide staining tank taken on the line 28-28 of Fig.
17, showing additional details of the tank interior.
Fig. 29 is a side sectional elevational view taken on the line 2~-29 of Fig. 19, showing details of the tank interior when in engagement with the loaded slide holder.
FigO 30 is a front sectional elevational view taken on the line 30-30 of Fig. 29, showing additional details of the tank interior when engaged with the loaded slide holder and showing the resultant upward displacement of a staining fluid about the slides.
Des~ri~tion of the Preferred Embodiments A conventional slide 20 is shown particularly in Figs. 2 and 4. The slide 20 is simply a rectangular piece of glass 21 on which a specimen 22 is mounted ~or microscopic examination. The slide has opposed flat sur-aces or faces 23 and 25, opposed long edges 26 and 27, and opposed short edges 28.
The most commonly used slide has a length (Ls) of approximately 3", a width (Ws) of approximately 1", and a thickness (Ts) of approximately lmm. In some instances, the slide may have a LS of 75mm and Ws of 25mm, or a TS
of 1.2mm.
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. 9 qlhese dimensions may vary slightly with different manufacturers. For instance, thicknesses may vary from .0525" to .0325~, and the width may vary from l.Q16" t~
.964".
The specimen 22 is prepared and then affixed to a face of the slide in any well known manner such as by use of air drying, heat, or reagent ~ixation. The specimen 22 is generally positioned at the center of one of the slide faces, with a clear zone, free of the specimen, on each end of the face. The specimen is virtually invisible at this point, prior to staining.
The slide 20, with the unstained specimen 22 thereon, is intended to be inserted into a disposable slide-staining device 30~ Device 30 is desirably formed of a transparent or translucent plastic into a generally long rectangular configuration having proportions yenerally conforming to the proportion of ~he slide 20. The device is intended to be used vertically, and has a top 31, a bottom 32, edges 33 and sides 35 and 36 A collar or flange 29 surrounds top 30 and has a flat surface thereon.
An internal cavity or chamber 40, likewise is rectangular in configuration and again, conforms generally to the proportions of the slide 20. The internal cavity 40 has opposed tapered vertical tracks 41 and 42, along the vertical edges 43 and 45 of the cavity extending from the top 40 of the cavity to the bottom 47. The cavity also has opposed sides 48 and 50.
The cavity 40 has at its upper end a sloping~
downwardly converging portion 51 having beveled edges 52 and sides 53.
-1,0-A cover or seal 55 is a~fixed to the top 31 on collar or flange 29 by suitable means, such as heat sealing or adhesive. The cover is intended to be peeled back by grasping and pulling on tab 56 prior to use, so that the cavity 40 is exposed A pool of staining liquid 60 is contained within the caYity and extends about halfwa~ up the cavity, as seen in Figs. 1 and 10.
The device is intended to be manufactured at a central facility whereat it is sealed with the staining liquid therein, as seen in ~ig. 1.
As seen in Figs. 5 through 8, there is shown in sequence views of a slide 20 being inserted into the cavity 40. In FigO 5, the slide 40 with a biological specimen 22 to be s~ained af~ixed thereon, is shown entering the cavity 40 on tracks 41 and 42. The slide 20 i5 spaced from the opposing sides 48 and 49 as seen best in Figs. lS and 16. The slide in Fig. 5 is just about to contact the upper surface of the pool 60, which is the static liquid level.
20 The static liquid level is at a heighth of Hl above the bottom of the cavity.
In Figs. 6 and 7, the slide progressively descends into the cavity below the pool, the liquid in the pool is displaced and rises until the dynamic fluid level reaches H2 as seen in Fig. 8, when the slide is fully inserted.
At this point, the staining liquid entirely covers specimen 22. Hl is approximately one~half of ~2. H2 also represents the heighth of the cavity as seen best in Fig. 14.
In Fig. 9~ the principle described and shown in Figs. 5 through 8 is illustrated in yraph form. As the slide is submerged below the static liquid level, there is a corresponding rise in the dynamic liquld level of the staining liquid in virtually a straight line relationship.
C~7~
As the slide is removed from the staining li~uid, after being left therein for an adequate period to accomplish staining, the slide is withdrawn and the reve~se s~quence occurs. The liquid level alls correspondingly as the liquid drains from the side of the slide into the pool. There is some residual staining liquid that adheres to the speci-men and the slide, and of c~urse the pool heighth at the end of staining drops correspondingly below the static liquid level, when the slide is withdrawn from the device. The slide when withdrawn from the device 30 has thereon the stained specimen 54 as seen in Fig. 4. Since the device is intended for a one-time use, such drop in the po~l heighth and volume is not objectionable.
The tapPred or guide portion 51 with beveled edges lS 52 and beveled side 53 permits easy insertion of the slide into the tracks. It also acts as a well to catch any over-flow of the staining liquid, including any splashing where the slide is dropped rapidly into the cavity. A collar or guide portion 29 extends around the upper end or top f the device and is integral therewith. Flange 29 has desirably a flat top portion 63 which permits a foil or cover 55 to be suitably secured thereto as by adhesive, or by a s~itable heat-sealing technique. The foil can be of a metallic or a plastic material capable of being peeled back from the covering by grasping a tab portion 56 and pulling to a completely open position, as seen in Fig. 3.
The cap portion in sealing and covering position is shown in FigsO 10 and 11.
There is variability in the width and thickness f slidesl even those claiming to have specific dimensions.
By tapering the tracks, the device will receive a slide of variable thickness and variable width and still keep the slide away from the inside wall of the cavity. Addi-tionally, the tapered tracks assure that only the edge of the slide will touch the track and not the surface which contains the specimen.
The slide 20 when inserted into the cavity 40 on ~he tracks 41 and 42 assumes a very definite position and posture within the cavity and witi- resp~ct to the sides or walls 48 and 49 of the cavity 40.
As seen in Fi~s. 14 to 16 inclusive, the slide 20 when inserted into its ~ottommost position has a top portion 61 which extends above the cavity 40 and top 31 of the device 30. The top portion ~1 of the slide 2D is clear of any specimen 22, since the specimen zone is conven-tionally in the middle of the slide as shown in Fig. 14 hy phantom lines. The portion 61 of the slide 20 remaining above the device 30 is used to grasp the slide for inserting into the device and for withdrawing the slide from the device.
The slide is also grasped by this portion during the rinsing subsequent to the staining. In this way, the operator's fingers remain free of contact with the staining solution, and free of the dye. Approximately one-sixth of the length of ~he slide LS extends above the device 30.
The spec;men 22 is stained when the slide is fully inserted as shown particularly in Figs. 14 through 16 inclu-sive. The slide as seen particularly in Figs. 15 and 16 i5 positioned centrally of the cavity 40 and spaced from walls or sides 48 and 49. The slide 20 has adjacent to each face thereof spaces 65 and 66 which contain staining liquid. The slide can be inserted with the specimen 22 facing in either direction as the slide 20 enters the cavity.
The face of the slide 20 and the specimen 22 thereon is positioned so that the specimen 22 cannot contact the sides o~ the cavity, but rather the specimen is held in a way that exposes it to the staining liquid in the space 65 or 66 formed ~y the slide and the side of the cavity. A minimum amount of stainin~ llquid is necessary to stain the specimen when the slide is so positioned. This is clearly shown in the sectional views in Figs. 15 and 16.
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~13-It will be seen from the above description that the space 65 anc7 66 adjacent to each face of the slide when the slide is fully inserted into the cavity is a flat thin space which contains a suitable amount of staining liq~id to properly stain the specimen thereon, without excess use of such liquid. ~rthermore, the slide itself is used to position this liq~id adjacent to the faces of the slide, and the specimen on one of the faces. It will be seen that it does not matter as to which direction the specimen ~aces when inserted into the cavity, since there is film on both faces of the slide.
The slide itself, in cooperation with the container, creates the film adjacent to the slide faces for staining, and then permits the liquid in those spaces to drain back lS into the bottom o~ the cavity and pool formed therein when the slide is removed.
In Figs. 10 throu~h 16 inclusive, there is shown in detail the interior of the device of Fig. 1. Ws is the width of the slide ~l LS is its length, and TS is its thick-ness~ ~2~ which is the heighth oE ~he cavity, as well asthe heighth of the dynamic liguid level discussed above, is approximately three~quarters of the slide len~th Ls~
The teacks 41 and 43 comprise tapered walls in the edges of the cavity, as shown. The width between the 2~ tracks, WT, as seen in Fig. 13! is sli~htly greater, for instance, .02~ greater, than the width of the slide Ws, so that there is adequate clearance for the slide to move in the tracks. TT, the thickness of the track at its tapered end, is slightly less, for instance, .016'i less, than the 30 thickness of the slide Ts~ Y, the thickness oF the cavity, is approximately twice the slide thickness, Ts. By virtue of the tapered tracks 41 and 42, the slide, when inserted în the tracks, is centr~lly spaced between the side walls ,~f;~ 7~
of the cavity, 48 and 49. Where the thickness of the cavity is twice that of the sli.de, the thickness of the spaces formed on each face of the slide when the slide is inserted will be approximately one-half the thickness o the slide.
By virtue of the tapered tracks 41 and 42, slides with slightly varying dimensions ~s set forth above will be suitably spaced within cavity 40. The tapered walls of the tracks 41 and 42 will compe~sate for these slight variations by guiding the slide at its edges thereof while still positioning the slide. Even with a slide having the smallest width andJor thickness within the variable dimen-sions, the slide is still spaced from the sides of the cavity, although all edges of the slide may not be in firm contact with the tracks. Even though such a relatively small slide ~ay have more freedom to move about within the cavity than a relatively large slide, the restraint imposed on the corners of the slide by the tapered walls of the tracks keeps the slide spaced from the cavity walls so that-a film of staining fluid can be formed adjacent the slide faces.
It should be understood that the dimensions given above are merely illustrative to indicate a suitable dimen-sioned container to practice the invention~
~eferring now to the preferred embodiment of the invention ~or simultaneously staining a multiplicity of slides, and with partic~lar reference to Figs. 17-30, slides 120 are of t.he nature, size and design described above with respect to slide 20.
The containe 130 may be genexally similar to the container 30 described above, in shape, heighth, and 30 width, except the horizontal depth of the container is ex- ~ :
tended to receive a plurality of slides, rather than onl~
one, as will be described. The container 130 has an open top 131, a bottom 132, edges 133, 134, sides 135, and 136, and collar or flange 129.
The internal cavity or main chamber 140 of con-tainer 130, has fo~r tracks s~ch as 141 and 142 in its four vertical corners, vertical edges 143 and 145, a bottom 1~7, and opposed sides 14B and 150.
In the version shown, the main chamber has at its top an overflow chamber 160 formed in the flange portion 129 which has side walls 168,170 formed by sides of the mai~ chamber 140, end walls 172,174 and bottom ledges such as 180.
~0 A seal cover 186, similar to seal 55 described above, is affixed to top 131 of the container.
The slides are spaced, positioned, and securely held in holder 200, which consists essentially of a drawer 202, a case 204, and spacers such as 206. The spacers are slidably and pivotally suspended in case 204 on pivot trun-nion pins such as 210 which ride in tracks 212 and 214 on the interiors of the opposed sidewalls 215,216 of the case.
Drawer 2~2 slides into case 204 so that when moved inwardly it pushes aqainst at least the foremost spacer 217~
The case 204 has a closed top wall 2~8 and an open bottom. (The case is shown inver~ed, in the loading position, in Figs. 20, 21, 22, 23 and 25. ) The side walls 215~216 contain respective slots 224 and 226, an open front and a closed back wall 230.
The tracks 212 and 214 in the case 204 are each formed by parallel ridges which deine channels 232, 234 for receiving the pins 210 on the spacers.
The spacers are thin, flat rectangles which pre-ferably have, at their long edges nearest the pins, a tapered edge or point for facilitating insertion between them of one end of the slides~ The pins are near the edges o~ the spacers so the spacers hang free and loose in the trAcks before loading of the slides. Preferably, these spacers are all alike.
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The drawer 202 has a front wall 300, push and pull knobs or ears 302, 304 at the opposite sides of the front wall, sides 306,308, and diagonal interior braces 310, 312 extending b~tween the front wall and the side walls
5 of the drawer~ Th~ side walls 306,308 each have a cam tooth such as 314 in~ended to ride in a corresponding slot such as 226 in the side wall of the case when the drawer is in-serted into the case. The drawer side walls fit inside of, and in sliding engagement with, the case side walls.
Each of the cam teeth 314 has a sloping surface such as 320 which permits the drawer to be inserted into the case until the teeth engage in their corresponding slots 224 and 226, after which the flat end surface 322 on the tooth prevents the drawer from uninter.tionally being pulled all the way 15 out of the case.
The drawer is desirably made of a relatively flexi-ble plastic materi~l such as ~olyethylene, wherein there is a degree of give~ par~icularly to the sides beyond the braces, for putposes which will be explained later. The 20 spacers are likewise desirably made of such flexible material.
In contrast to the drawer and space~s, the case is made of a relatively rigid material such as glass-filled nylon, so that the relatively flexible drawer and spacers, in com-bination with the relatively rigid case, will permit the 25 drawer to lock the spacer~ to the case as described herein-after.
It is intended that container 130 contain a con-fined or limited pool of s~aining fluid 360. As best seen in Figures 27 and 2~, this pool extends to a height H~ which 3~ is preferably somewhat over one-half of the height of the chamber 140.
As with the single slide embodiment shown in Figs.
1 through 16, it is intended that slides 120 be inserted downwardly into this pool 360, whereby staining liquid o the pool will be displaced upwardly in the main chamber 5 substantially to the top thereof without overflowing into the overflow chamber, but in the event of excess upward flow of displaced liquid this may be caught in the over~
flow chamber 160. In a preferred design, the proportion of parts and depth of pool are such that no substantial 10 overflow occurs, and in such case the large overflow cha~ber can be eliminated and the top of the container made as in the single-slide version of Figs. 1-46.
The slides are held in exact spaced relationship with each other and the container so that a thin film is 15 formed on both sides of the slides. The space between the slides conforms to the thickness of the spacers. Desirably, this thickness is approximately equal to the thickness of the slides themselves, which as indicated above can be about 1 mm. The spacing is such that when the spaced slides are 20 secured in the holder, and the slides are immersed in the limited pool in the container substantially to ~he bottoM
thereof, the pool will be displaced so that the staining liquid rises in thin films which reach and cover substan-tially all the slide portion which extends into main chamber 25 of the container, and particularly the portions to which the specimens are affix~d.
The positioning and spacing of the slides are accomplished in the holder, which initially is placed on its top, in an inverted position. The spacers are allowed 30 to hang freely on their trunnion pins in tracks 212,214 as seen in Figs. 22, 23 and 240 At this tîme, the drawer 202 ;
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is open, and side walls 306 and 308 of the drawer are para-llel ~n~ in a rela~ed ~osition, whereb~ the dra~er can slide Freely in the c~se, except that the drawee cannot be with-drawn compLetely out beca~se of the interfering action oE
cam teeth 314 As shown in Fig. 22, slides 120 are then inserted between the spacers such as 206, with the beveled edges of the spacers aiding entry of the slides between the spacers.
The spacers and slides are preferably of the same width.
The width of the case between its side walls is equal to the width of the slides plus a slight clearance dimension.
During assembly, the slides are dropped to the "bottom"
of the inverted case, so that the straight edges of the ends of the slides rest on the interior of the top wall of the case. When the necessary n~mber of slides has been inserted in the holder, which is five for the five-slide holder and container embodiment shown, the drawer is pushed closed from the position shown in Figs. 23 and 24 to the position shown in Figs. ~5 and 26.
Sidewalls 306 and 308 of drawer 202 terminate at their free ends in respective wedges 370, 372 having respective, confronting, tapered wedging surfaces 374,376 adapted to enter the above-mentioned clearances between ~pacers and side walls when the drawe~ is closed. Also, when tl,e drawer is closed, the outer ends of sidewalls 306 and 308 are spread outward due to the wedging actions of surfaces 374, 376 against the ends of the spacers. The wedges are formed in the side walls beyond the braces 310, 312, so that the side walls are held relatively parallel up to the end of the braces, while the remaining outer end portions of the side walls are relatively free to flex out-ward under the influence of the spacers, and against the inside of the side wal]s of the case. The resistance of the spacers and slides against the wedges therefore serves to jam the wedges against the side walls of the drawer, ('7~
so that the drawer, the slides and the spacers are locked in position, with the slides clamped between the spacers.
The spacers, Which are relatively flexible, give slightly against ~he har~ glass of the slides and then serve to cushion-grip the slides. ~he slides are now held parallel to one another, and evenly spaced from each other by pre-determined distances.
If there is any slight misalignment of the slides because the ends of the slides are not touching the interior of the case top, or because the slide ends are not entirely square with the slide sides, any such slight misalignment can be remedied by forcing the slides into the correct posi-tion manually, with one's fingers.
The outside dimensions of the five-slide group extending from the holder closely conform to the chamber designed for the ive slide staining operations~ These dimensions are such that when the holder is inverted from the loading position and the exposed portions of the slides inserted into the container, the edges of the outside slides touch the sides of the tracks such as 141,142 in the con-tainer, whereby, when the slides, by means of the holder, are inserted into the container, there is formed a thin space adjacent to each of the outside faces of the outside slides. The tracks such as 1~1,142 can be molded integrally with the remainder o~ the container, and may have lead-in bevels at their entrance ends to facilitate insertion of the slide assembly.
As the slides are immersed into the pool, the liquid is displaced upwardly along the slide faces. The dimension of ~he portion of the slides extending into the holder is preferably such that when the holder comes into contact with the top of the container, the slides almost (i 7 ~ ~
touch, or are just slightly above, the bottom of the con-tainer. Thus~ essentially all of the staining liquid at this time extends in thin films across the faces of the ~lides, and across the speci~ens on the slides. The liquid rise~ to the top of the chamber, and permis~ibly int~ the overflow chamber if one or more of the slides is slightly thicker than a generally standard slide.
When the specimen~ have been thus stained, the slides are withdrawn from the container by means of the holder. The staining fluid drains off the slides, back into a pool of approximately the same size as the beginning pool minus the quantity which adheres to the slide. The container can permissibly be resealed with the original seal, and then neatly discarded. The holder and slides are then inverted and the slides unloaded from the holder in essentially a reverse operation from the loading opera-tion described above.
In order to withdraw the drawer from the case and to release the grip of the spacers on the slides, it is necessary to break the wedging action of the sides of the deawer. To do this, the drawer is gripped at its ears 302 and 304 between the fingers, and an alternating rocking motion is exerted on the drawer as the ~rawer is withdrawn from the case, which is anchored by the fingers of the operator's other hand. The case is desirably resting on a table during the unloading operation. After the jamming and wedging action of the drawer is thu~ broken, the dr~wer is withdrawn to release the spacers and ~lides to the position shown in Figs. 23 and 24. The slides can then be delicately re-moved from the holder and further processed by rinsing andthe like, in the usual wayO
The holder carl optionally be washed and rinsed and then used with a new container and pool, or the holder can be thrown away.
It is intended that the container and holder be mass-produced out of plastic, by, for instance, extr~sion molding, so that the cost per unit be relatively low.
The above embodiment is illustrated with a five-slide group. It should be understood that the same principlecan be used with respect to any desired group from, for instance, two to ten and beyond, with each having its speci-fically dimensioned container, holder, and pool. However, it is contemplated that the five-slide arrangements will be the most prevalent.
Two such five-slide holders can also be clipped back-to-back to create a ten-slide holder. Such ten-slide combination may then be inserted into a ten-slide container, designed in accordance with the invention to accommodate the ten slides.
The container shown above has an overflow compart-ment as described~ This is generally desirable when it is anticipated that the slides used vary somewhat in their thickness. Since a thicker slide displaces more liquid, the over~low chamber serves to receive and hold this over-flow during the staining operation. Where the slides are relatively uniform in thickness, there is no need for an overflow chamber, since the displacement is more controlled.
In this condition, the overflow chamber can be reduced in sizep or eliminated, as seen in the embodiment shown in Figs~ 1 to 16.
As an aid in design, the following relations are noted. The distance H2 by which the level of the top of the pool of staining liquid is raised by immersion of the slides is equal to the volume of the liquid displaced upward-ly by the glass of the slides, divided by the average cross-sectional area of the glass-free chamber space into which the liquid is displaced. Designating the distance by which the slides are immersed below the top of the original pool as ~1~ the average cross-sectional area of the chamber above D ( r~7~
the original pool surface with the slides removed as ACh, and the average cross-sectional area of slide glass in the liquid as Agl, then H2 =
ch gl In the simple case of a purely reetangular chamber, ~ive slides of l mm thickness each spaced from each other and from the chamber wal~s by l mm, and immersion of the slides to the full depth H of the original pool, the formula pre-diets a rise in liquid level H2 = H = H = 83H
In praetice, the height H2 will be redueed below ~his valueby any extra ehamber spaee at the edges of the inserted slides, and inereased due to reduetions in ehamber space eaused by tracks in the chamber.
While the invention has been described with par-tieular referenee to specific embodiments i.n the interest of eomplete definiteness, it will be understood that it may be embodied in a variety of ~orms diverse from those speeifieally shown and described, without departing from the spirit and seope of the invention as defined by the appended elaims.
Each of the cam teeth 314 has a sloping surface such as 320 which permits the drawer to be inserted into the case until the teeth engage in their corresponding slots 224 and 226, after which the flat end surface 322 on the tooth prevents the drawer from uninter.tionally being pulled all the way 15 out of the case.
The drawer is desirably made of a relatively flexi-ble plastic materi~l such as ~olyethylene, wherein there is a degree of give~ par~icularly to the sides beyond the braces, for putposes which will be explained later. The 20 spacers are likewise desirably made of such flexible material.
In contrast to the drawer and space~s, the case is made of a relatively rigid material such as glass-filled nylon, so that the relatively flexible drawer and spacers, in com-bination with the relatively rigid case, will permit the 25 drawer to lock the spacer~ to the case as described herein-after.
It is intended that container 130 contain a con-fined or limited pool of s~aining fluid 360. As best seen in Figures 27 and 2~, this pool extends to a height H~ which 3~ is preferably somewhat over one-half of the height of the chamber 140.
As with the single slide embodiment shown in Figs.
1 through 16, it is intended that slides 120 be inserted downwardly into this pool 360, whereby staining liquid o the pool will be displaced upwardly in the main chamber 5 substantially to the top thereof without overflowing into the overflow chamber, but in the event of excess upward flow of displaced liquid this may be caught in the over~
flow chamber 160. In a preferred design, the proportion of parts and depth of pool are such that no substantial 10 overflow occurs, and in such case the large overflow cha~ber can be eliminated and the top of the container made as in the single-slide version of Figs. 1-46.
The slides are held in exact spaced relationship with each other and the container so that a thin film is 15 formed on both sides of the slides. The space between the slides conforms to the thickness of the spacers. Desirably, this thickness is approximately equal to the thickness of the slides themselves, which as indicated above can be about 1 mm. The spacing is such that when the spaced slides are 20 secured in the holder, and the slides are immersed in the limited pool in the container substantially to ~he bottoM
thereof, the pool will be displaced so that the staining liquid rises in thin films which reach and cover substan-tially all the slide portion which extends into main chamber 25 of the container, and particularly the portions to which the specimens are affix~d.
The positioning and spacing of the slides are accomplished in the holder, which initially is placed on its top, in an inverted position. The spacers are allowed 30 to hang freely on their trunnion pins in tracks 212,214 as seen in Figs. 22, 23 and 240 At this tîme, the drawer 202 ;
(77~
is open, and side walls 306 and 308 of the drawer are para-llel ~n~ in a rela~ed ~osition, whereb~ the dra~er can slide Freely in the c~se, except that the drawee cannot be with-drawn compLetely out beca~se of the interfering action oE
cam teeth 314 As shown in Fig. 22, slides 120 are then inserted between the spacers such as 206, with the beveled edges of the spacers aiding entry of the slides between the spacers.
The spacers and slides are preferably of the same width.
The width of the case between its side walls is equal to the width of the slides plus a slight clearance dimension.
During assembly, the slides are dropped to the "bottom"
of the inverted case, so that the straight edges of the ends of the slides rest on the interior of the top wall of the case. When the necessary n~mber of slides has been inserted in the holder, which is five for the five-slide holder and container embodiment shown, the drawer is pushed closed from the position shown in Figs. 23 and 24 to the position shown in Figs. ~5 and 26.
Sidewalls 306 and 308 of drawer 202 terminate at their free ends in respective wedges 370, 372 having respective, confronting, tapered wedging surfaces 374,376 adapted to enter the above-mentioned clearances between ~pacers and side walls when the drawe~ is closed. Also, when tl,e drawer is closed, the outer ends of sidewalls 306 and 308 are spread outward due to the wedging actions of surfaces 374, 376 against the ends of the spacers. The wedges are formed in the side walls beyond the braces 310, 312, so that the side walls are held relatively parallel up to the end of the braces, while the remaining outer end portions of the side walls are relatively free to flex out-ward under the influence of the spacers, and against the inside of the side wal]s of the case. The resistance of the spacers and slides against the wedges therefore serves to jam the wedges against the side walls of the drawer, ('7~
so that the drawer, the slides and the spacers are locked in position, with the slides clamped between the spacers.
The spacers, Which are relatively flexible, give slightly against ~he har~ glass of the slides and then serve to cushion-grip the slides. ~he slides are now held parallel to one another, and evenly spaced from each other by pre-determined distances.
If there is any slight misalignment of the slides because the ends of the slides are not touching the interior of the case top, or because the slide ends are not entirely square with the slide sides, any such slight misalignment can be remedied by forcing the slides into the correct posi-tion manually, with one's fingers.
The outside dimensions of the five-slide group extending from the holder closely conform to the chamber designed for the ive slide staining operations~ These dimensions are such that when the holder is inverted from the loading position and the exposed portions of the slides inserted into the container, the edges of the outside slides touch the sides of the tracks such as 141,142 in the con-tainer, whereby, when the slides, by means of the holder, are inserted into the container, there is formed a thin space adjacent to each of the outside faces of the outside slides. The tracks such as 1~1,142 can be molded integrally with the remainder o~ the container, and may have lead-in bevels at their entrance ends to facilitate insertion of the slide assembly.
As the slides are immersed into the pool, the liquid is displaced upwardly along the slide faces. The dimension of ~he portion of the slides extending into the holder is preferably such that when the holder comes into contact with the top of the container, the slides almost (i 7 ~ ~
touch, or are just slightly above, the bottom of the con-tainer. Thus~ essentially all of the staining liquid at this time extends in thin films across the faces of the ~lides, and across the speci~ens on the slides. The liquid rise~ to the top of the chamber, and permis~ibly int~ the overflow chamber if one or more of the slides is slightly thicker than a generally standard slide.
When the specimen~ have been thus stained, the slides are withdrawn from the container by means of the holder. The staining fluid drains off the slides, back into a pool of approximately the same size as the beginning pool minus the quantity which adheres to the slide. The container can permissibly be resealed with the original seal, and then neatly discarded. The holder and slides are then inverted and the slides unloaded from the holder in essentially a reverse operation from the loading opera-tion described above.
In order to withdraw the drawer from the case and to release the grip of the spacers on the slides, it is necessary to break the wedging action of the sides of the deawer. To do this, the drawer is gripped at its ears 302 and 304 between the fingers, and an alternating rocking motion is exerted on the drawer as the ~rawer is withdrawn from the case, which is anchored by the fingers of the operator's other hand. The case is desirably resting on a table during the unloading operation. After the jamming and wedging action of the drawer is thu~ broken, the dr~wer is withdrawn to release the spacers and ~lides to the position shown in Figs. 23 and 24. The slides can then be delicately re-moved from the holder and further processed by rinsing andthe like, in the usual wayO
The holder carl optionally be washed and rinsed and then used with a new container and pool, or the holder can be thrown away.
It is intended that the container and holder be mass-produced out of plastic, by, for instance, extr~sion molding, so that the cost per unit be relatively low.
The above embodiment is illustrated with a five-slide group. It should be understood that the same principlecan be used with respect to any desired group from, for instance, two to ten and beyond, with each having its speci-fically dimensioned container, holder, and pool. However, it is contemplated that the five-slide arrangements will be the most prevalent.
Two such five-slide holders can also be clipped back-to-back to create a ten-slide holder. Such ten-slide combination may then be inserted into a ten-slide container, designed in accordance with the invention to accommodate the ten slides.
The container shown above has an overflow compart-ment as described~ This is generally desirable when it is anticipated that the slides used vary somewhat in their thickness. Since a thicker slide displaces more liquid, the over~low chamber serves to receive and hold this over-flow during the staining operation. Where the slides are relatively uniform in thickness, there is no need for an overflow chamber, since the displacement is more controlled.
In this condition, the overflow chamber can be reduced in sizep or eliminated, as seen in the embodiment shown in Figs~ 1 to 16.
As an aid in design, the following relations are noted. The distance H2 by which the level of the top of the pool of staining liquid is raised by immersion of the slides is equal to the volume of the liquid displaced upward-ly by the glass of the slides, divided by the average cross-sectional area of the glass-free chamber space into which the liquid is displaced. Designating the distance by which the slides are immersed below the top of the original pool as ~1~ the average cross-sectional area of the chamber above D ( r~7~
the original pool surface with the slides removed as ACh, and the average cross-sectional area of slide glass in the liquid as Agl, then H2 =
ch gl In the simple case of a purely reetangular chamber, ~ive slides of l mm thickness each spaced from each other and from the chamber wal~s by l mm, and immersion of the slides to the full depth H of the original pool, the formula pre-diets a rise in liquid level H2 = H = H = 83H
In praetice, the height H2 will be redueed below ~his valueby any extra ehamber spaee at the edges of the inserted slides, and inereased due to reduetions in ehamber space eaused by tracks in the chamber.
While the invention has been described with par-tieular referenee to specific embodiments i.n the interest of eomplete definiteness, it will be understood that it may be embodied in a variety of ~orms diverse from those speeifieally shown and described, without departing from the spirit and seope of the invention as defined by the appended elaims.
Claims (17)
1. A system for staining specimens on a plurality of microscope slides, each of said specimens extending by no more than a distance D from one end of its associated slide, comprising:
a) a slide holder for holding the other ends of said slides in a clamped assembly with their lower portions extending from said holder in parallel confronting relation to each other;
b) a container having a main chamber partially filled with staining liquid, the depth of said liquid in said chamber being less than said distance D;
c) said slides in said slide holder being suf-ficiently close together, and the walls of said main chamber conforming sufficiently closely to the periphery of said lower portions of said slides when said assembly is inserted downwardly into said pool, that said staining liquid rises in said chamber to cover said specimens.
a) a slide holder for holding the other ends of said slides in a clamped assembly with their lower portions extending from said holder in parallel confronting relation to each other;
b) a container having a main chamber partially filled with staining liquid, the depth of said liquid in said chamber being less than said distance D;
c) said slides in said slide holder being suf-ficiently close together, and the walls of said main chamber conforming sufficiently closely to the periphery of said lower portions of said slides when said assembly is inserted downwardly into said pool, that said staining liquid rises in said chamber to cover said specimens.
2. The apparatus of Claim 1, wherein said con-tainer comprises guide means for holding the outer faces of the end slides in said assembly spaced from the walls of said chamber.
3. The apparatus of Claim 2, wherein said main chamber has a generally rectangular cross-section and said guide means comprises four vertical walls extending across the corners of said chamber for contacting the four corners of said end slides guidingly.
4. The system of Claim 1, wherein said slide holder comprises:
a case having a pair of side walls, a top wall, a back wall, an open bottom and an open front; and a clamp-ing drawer adapted to slide in and out of the open front of said case;
said case containing a plurality of slide spacers for receiving said slides between them and having track means in its opposed side walls slidably mounting said slide spacers for motion toward and away from said back wall;
said clamping drawer having spacer contacting means formed integrally with said side walls for compacting said spacers when said drawer is advanced in its closing direction, thereby to clamp said slides between said spacers.
a case having a pair of side walls, a top wall, a back wall, an open bottom and an open front; and a clamp-ing drawer adapted to slide in and out of the open front of said case;
said case containing a plurality of slide spacers for receiving said slides between them and having track means in its opposed side walls slidably mounting said slide spacers for motion toward and away from said back wall;
said clamping drawer having spacer contacting means formed integrally with said side walls for compacting said spacers when said drawer is advanced in its closing direction, thereby to clamp said slides between said spacers.
5. The system of Claim 4, wherein said spacer contacting means comprises wedge means responsive to motion of said drawer in its closing direction to be wedgingly inserted between at least one of said spacers and at least one adjacent wall of said case, thereby to lock said slides and said spacers releasably in said clamped assembly.
6. The system of Claim 5, wherein: each of said spacers comprises a pair of trunnions extending from op-posite sides thereof and into said track means; wherein each of said spacers is narrower than the distance between said side walls, so as to leave openings at each end of the spacer between spacer and adjacent side wall; and where-in said wedge means comprises a pair of forwardly-extending wedges, one adjacent each of said side walls, and aligned with said openings between said spacers and said side walls.
7. The system of Claim 6, wherein said container comprises an overflow chamber of greater horizontal cross-section than the main chamber for receiving excess displaced liquid from said pool.
8. A slide holder for releasably clamping a plu-rality of microscope slides in a temporary assembly in which said slides are disposed parallel to and aligned with each other, with their major faces in closely-confronting spaced-apart relation, comprising:
a case having opposed side walls spaced apart a distance greater than the widths of said slides, an open bottom, an open front, and track means extending along the inner sides of said side walls adjacent the bottom of said case;
a plurality of vertically-disposed spacers having track-engaging means slidingly cooperating with said track means, whereby said spacers can be spaced apart to receive slides between them or compacted to clamp said slides between them, by sliding said spacers along said track means in one direction or the other;
a drawer fitting slidingly into said case, said drawer having a front wall and a pair of opposed side walls terminating in a corresponding pair of tapered wedges having sloped, confronting, wedge surfaces;
said spacers having a width less than the interior width of said case thereby to provide spaces between at least the foremost of said spacers and the adjacent side walls of said case, into which spaces said wedges can enter wedgingly when said drawer is pushed inwardly, thereby to compact said spacers and clamp said slides between them;
said sidewalls of said drawer being sufficiently flexible to permit said wedges to be bent outwardly against said side walls of said case when said drawer is closed, thereby to lock said drawer temporarily in fixed position with respect to said case.
a case having opposed side walls spaced apart a distance greater than the widths of said slides, an open bottom, an open front, and track means extending along the inner sides of said side walls adjacent the bottom of said case;
a plurality of vertically-disposed spacers having track-engaging means slidingly cooperating with said track means, whereby said spacers can be spaced apart to receive slides between them or compacted to clamp said slides between them, by sliding said spacers along said track means in one direction or the other;
a drawer fitting slidingly into said case, said drawer having a front wall and a pair of opposed side walls terminating in a corresponding pair of tapered wedges having sloped, confronting, wedge surfaces;
said spacers having a width less than the interior width of said case thereby to provide spaces between at least the foremost of said spacers and the adjacent side walls of said case, into which spaces said wedges can enter wedgingly when said drawer is pushed inwardly, thereby to compact said spacers and clamp said slides between them;
said sidewalls of said drawer being sufficiently flexible to permit said wedges to be bent outwardly against said side walls of said case when said drawer is closed, thereby to lock said drawer temporarily in fixed position with respect to said case.
9. The slide holder of Claim 8, wherein said drawer comprises braces extending between the front wall of said drawer and positions on said side walls thereof short of said wedges, whereby portions of said lastnamed side walls short of said wedges are held rigid while said wedges are free to be bent outwardly during said locking action thereof.
10. The method of staining specimens on a multi-plicity of microscope slides, each of said specimens extend-ing by no more than a distance D from one end of its as-sociated slide, said method comprising:
forming a temporary assembly of said slides by clamping said slides together, with thin spacers between the end portions of said slides opposite from said one ends thereof, and with the remaining portions of said slides extending beyond said spacers in parallel, closely spaced-apart confronting relation to each other;
with said one ends of said slides positioned down-wardly, moving said temporarily-assembled slides downwardly to insert said remaining portions of said slides into a pool of staining liquid confined in a chamber having a depth prior to said inserting which is less than said distance D and having a horizontal cross-section fitting sufficiently closely about said inserted assembly that said liquid is displaced upwardly to cover all of said specimens with said staining liquid and thereby stain them; and thereafter lifting said temporary assembly to remove said slides from said liquid, and unclamping said stained slides to permit their removal from said assembly.
forming a temporary assembly of said slides by clamping said slides together, with thin spacers between the end portions of said slides opposite from said one ends thereof, and with the remaining portions of said slides extending beyond said spacers in parallel, closely spaced-apart confronting relation to each other;
with said one ends of said slides positioned down-wardly, moving said temporarily-assembled slides downwardly to insert said remaining portions of said slides into a pool of staining liquid confined in a chamber having a depth prior to said inserting which is less than said distance D and having a horizontal cross-section fitting sufficiently closely about said inserted assembly that said liquid is displaced upwardly to cover all of said specimens with said staining liquid and thereby stain them; and thereafter lifting said temporary assembly to remove said slides from said liquid, and unclamping said stained slides to permit their removal from said assembly.
11. The method of claim 10, comprising guiding opposite edges of at least one of said slides during said inserting and removal of said assembly to prevent contact between the walls of said chamber and the faces of the end slides in said assembly.
12. The method of claim 10, wherein said clamping of said slides comprises loosely supporting said spacers parallel to each other in a slide holder, inserting said end portions of said slides between said spacers, compacting said slides and spacers, and inserting wedge means between said compacted assembly and said slide holder; and wherein said unclamping comprises loosening said wedge means from between said compacted assembly and said slide holder.
13. A disposable staining device intended for one-time use for applying a staining liquid to specimens fixed to specimen zones on the faces of a plurality of glass slides, the specimen zone on each slide face being disposed substantially equidistant the slide ends, said device comprising:
a container having an upwardly opening cavity therein for receiving the slides;
a quantity of staining liquid partially filling said cavity; and a removable seal closing said cavity to contain said liquid prior to use of said device;
said cavity having a height shorter than the length of the slides to permit the ends of the slides to extend above the device when the slides are fully inserted in the cavity;
the specimen zone of each slide being disposed completely within the cavity when the slide is fully inserted into the cavity regardless of which end of the slide is so inserted;
means for maintaining the slides in a spaced parallel relationship and for preventing contact of the opposed faces of the specimen zone of the slide with the cavity walls when the slides are inserted in the cavity;
the depth of the staining liquid prior to insertion of the slides being less than the distance between the upper edge of the specimen zone of each slide and the inserted end of the slide;
the insertion of the slides displacing said liquid and raising the liquid level to a level sufficient to cover the specimen zones and thereby using a relatively minimum amount of staining fluid to stain the specimens fixed thereto.
a container having an upwardly opening cavity therein for receiving the slides;
a quantity of staining liquid partially filling said cavity; and a removable seal closing said cavity to contain said liquid prior to use of said device;
said cavity having a height shorter than the length of the slides to permit the ends of the slides to extend above the device when the slides are fully inserted in the cavity;
the specimen zone of each slide being disposed completely within the cavity when the slide is fully inserted into the cavity regardless of which end of the slide is so inserted;
means for maintaining the slides in a spaced parallel relationship and for preventing contact of the opposed faces of the specimen zone of the slide with the cavity walls when the slides are inserted in the cavity;
the depth of the staining liquid prior to insertion of the slides being less than the distance between the upper edge of the specimen zone of each slide and the inserted end of the slide;
the insertion of the slides displacing said liquid and raising the liquid level to a level sufficient to cover the specimen zones and thereby using a relatively minimum amount of staining fluid to stain the specimens fixed thereto.
14. The invention as claimed in claim 13 wherein the liquid substantially fills the cavity when the slides are inserted therein.
15. A method of staining specimens fixed to specimen zones on the faces of a plurality of glass slides, each specimen zone being generally centrally disposed on a slide face intermediate the slide ends, comprising the steps of:
providing a sealed disposable staining container intended for one-time use having a vertically opening cavity therein dimensioned to receive a major portion of each of a plurality of slides with the ends of the slides extending above the container when the slides are inserted substantially to the bottom of said cavity;
providing a quantity of staining liquid in said cavity having a static liquid level therein in the absence of inserted slides which is less than the distance between an end of a slide and the distant edge of the slide specimen zone;
providing means for maintaining the slides in closely-spaced parallel relation during insertion in the cavity and for preventing contact of the opposed faces of the specimen zone of the slide with the cavity walls;
removing the seal from the container;
inserting the slides into the cavity and thereby upwardly displacing the staining liquid from the static level to a dynamic level sufficient to cover the specimen zones of the slides and thereby stain the specimens fixed thereto when a slide is so positioned between the cavity walls;
removing the stained slides from the container;
and discarding the seal, container and staining liquid.
providing a sealed disposable staining container intended for one-time use having a vertically opening cavity therein dimensioned to receive a major portion of each of a plurality of slides with the ends of the slides extending above the container when the slides are inserted substantially to the bottom of said cavity;
providing a quantity of staining liquid in said cavity having a static liquid level therein in the absence of inserted slides which is less than the distance between an end of a slide and the distant edge of the slide specimen zone;
providing means for maintaining the slides in closely-spaced parallel relation during insertion in the cavity and for preventing contact of the opposed faces of the specimen zone of the slide with the cavity walls;
removing the seal from the container;
inserting the slides into the cavity and thereby upwardly displacing the staining liquid from the static level to a dynamic level sufficient to cover the specimen zones of the slides and thereby stain the specimens fixed thereto when a slide is so positioned between the cavity walls;
removing the stained slides from the container;
and discarding the seal, container and staining liquid.
16. The invention as claimed in claim 15 including the step of resealing the container cavity prior to discarding the container, thereby maintaining the staining liquid within the container cavity.
17. The invention as claimed in claim 15 wherein the dynamic liquid level is substantially at the top of the cavity.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US46251083A | 1983-01-31 | 1983-01-31 | |
| US462,510 | 1983-01-31 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CA1200744A true CA1200744A (en) | 1986-02-18 |
Family
ID=23836703
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA000426723A Expired CA1200744A (en) | 1983-01-31 | 1983-04-26 | Method and structure for staining of biologic slides |
Country Status (2)
| Country | Link |
|---|---|
| JP (1) | JPS59142436A (en) |
| CA (1) | CA1200744A (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US7588733B2 (en) * | 2003-12-04 | 2009-09-15 | Idexx Laboratories, Inc. | Retaining clip for reagent test slides |
-
1983
- 1983-04-26 JP JP58072315A patent/JPS59142436A/en active Pending
- 1983-04-26 CA CA000426723A patent/CA1200744A/en not_active Expired
Also Published As
| Publication number | Publication date |
|---|---|
| JPS59142436A (en) | 1984-08-15 |
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