US2899363A - Treatment of microscope specimen slides - Google Patents
Treatment of microscope specimen slides Download PDFInfo
- Publication number
- US2899363A US2899363A US2899363DA US2899363A US 2899363 A US2899363 A US 2899363A US 2899363D A US2899363D A US 2899363DA US 2899363 A US2899363 A US 2899363A
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- United States
- Prior art keywords
- slide
- solution
- cells
- ether
- specimen
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- Expired - Lifetime
Links
- 239000000463 material Substances 0.000 claims description 21
- 239000002904 solvent Substances 0.000 claims description 18
- 238000000034 method Methods 0.000 claims description 15
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 14
- 229910001868 water Inorganic materials 0.000 claims description 14
- POAOYUHQDCAZBD-UHFFFAOYSA-N 2-butoxyethanol Chemical compound CCCCOCCO POAOYUHQDCAZBD-UHFFFAOYSA-N 0.000 claims description 9
- 239000007788 liquid Substances 0.000 claims description 8
- 238000010186 staining Methods 0.000 claims description 7
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 28
- 210000004027 cell Anatomy 0.000 description 22
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 11
- 239000000203 mixture Substances 0.000 description 11
- 229920003086 cellulose ether Polymers 0.000 description 10
- 235000019441 ethanol Nutrition 0.000 description 7
- 239000001866 hydroxypropyl methyl cellulose Substances 0.000 description 6
- 229920003088 hydroxypropyl methyl cellulose Polymers 0.000 description 6
- 235000010979 hydroxypropyl methyl cellulose Nutrition 0.000 description 6
- UFVKGYZPFZQRLF-UHFFFAOYSA-N hydroxypropyl methyl cellulose Chemical compound OC1C(O)C(OC)OC(CO)C1OC1C(O)C(O)C(OC2C(C(O)C(OC3C(C(O)C(O)C(CO)O3)O)C(CO)O2)O)C(CO)O1 UFVKGYZPFZQRLF-UHFFFAOYSA-N 0.000 description 6
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 4
- 229920013820 alkyl cellulose Polymers 0.000 description 3
- 239000002775 capsule Substances 0.000 description 3
- 230000006378 damage Effects 0.000 description 3
- 238000006703 hydration reaction Methods 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 230000002496 gastric effect Effects 0.000 description 2
- 235000011187 glycerol Nutrition 0.000 description 2
- 239000004094 surface-active agent Substances 0.000 description 2
- 229920002994 synthetic fiber Polymers 0.000 description 2
- 108010077544 Chromatin Proteins 0.000 description 1
- FBPFZTCFMRRESA-FSIIMWSLSA-N D-Glucitol Natural products OC[C@H](O)[C@H](O)[C@@H](O)[C@H](O)CO FBPFZTCFMRRESA-FSIIMWSLSA-N 0.000 description 1
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 description 1
- FBPFZTCFMRRESA-JGWLITMVSA-N D-glucitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-JGWLITMVSA-N 0.000 description 1
- 229930195725 Mannitol Natural products 0.000 description 1
- 239000003708 ampul Substances 0.000 description 1
- 210000000481 breast Anatomy 0.000 description 1
- 125000004432 carbon atom Chemical group C* 0.000 description 1
- 229920002678 cellulose Polymers 0.000 description 1
- 239000001913 cellulose Substances 0.000 description 1
- 210000003483 chromatin Anatomy 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 230000018044 dehydration Effects 0.000 description 1
- 238000006297 dehydration reaction Methods 0.000 description 1
- 235000014113 dietary fatty acids Nutrition 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 230000002183 duodenal effect Effects 0.000 description 1
- 230000002357 endometrial effect Effects 0.000 description 1
- 230000006862 enzymatic digestion Effects 0.000 description 1
- 150000002148 esters Chemical class 0.000 description 1
- 238000001704 evaporation Methods 0.000 description 1
- 230000008020 evaporation Effects 0.000 description 1
- 239000000194 fatty acid Substances 0.000 description 1
- 229930195729 fatty acid Natural products 0.000 description 1
- 150000004665 fatty acids Chemical class 0.000 description 1
- 239000000594 mannitol Substances 0.000 description 1
- 235000010355 mannitol Nutrition 0.000 description 1
- 238000012856 packing Methods 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 230000001681 protective effect Effects 0.000 description 1
- 239000000600 sorbitol Substances 0.000 description 1
- 210000002700 urine Anatomy 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/30—Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
Definitions
- the present invention is directed to the subject of microscopic examination of body cells and more particularly to a method of treating the transparent slides in order to preserve the specimens for such examination.
- compositions for carrying out the method which composition is stable, is easily handled and is highly effective in its application to the slide.
- a solution of a water-soluble synthetic film-forming material which preferably also has the property of reducing surface tension.
- the solvent used is generally a mixture of ethyl ether and ethyl alcohol.
- the specimen smear on the slide is treated with the solution by flowing said solution onto the slide to cover the specimen. It is important that the slide be not dipped into the solution as otherwise at least part of the cells of the specimen may be washed off.
- the solvent is allowed to evaporate, thereby not only fixing the cells but also forming a film thereon to protect it during transit.
- the slide is then treated with an aqueous ether alcohol liquid which dissolves the film.
- an alkyl cellulose ether soluble in water is an ideal substance for the purpose. Specifically there is provided for immediate fixation and surface coating a capsule containing .2 gram of hydroxy propyl methyl cellulose ether, and dissolving the cellulose ether in 10 cc. of water at a temperature of, say, 80 to 90 C. To this solution is added 190 cc. of a mixture of in 3 parts of water to 2 parts of a mixture of equal equal parts of ethyl alcohol and ethyl ether.
- the cellulose ether preferably has a viscosity of about 4,000 centipoises although cellulose ethers of other viscosities are also usable.
- a cervical or a vaginal material is placed upon the slide.
- the specimen may be allowed to dry in the air.
- the slide is treated with the use of a solution of a surface active agent in water in a suitable concentration. It has been found that the sorbitol or mannitol partial esters of higher fatty acids such as those having 16 and 18 carbon atoms are suitable for the purpose.
- the specimen is then fixed, stained and examined microscopically.
- the capsule containing .2 g. hydroxy propyl methyl cellulose ether has the property of reducing the surface tension of water to about 43 dynes per centimeter, in a solution of 400 cc. of water and 200 cc. of a mixture of 95% ethyl alcohol and ethyl ether. Dried and unfixed slides are dipped in this solution for about five minutes, fixed in ether-alcohol for about ten minutes and then stained.
- the flowing of the cellulose ether solution onto the slide prevents the washing away of cells and after drying the specimen may be handled even carelessly without damage to the specimen.
- the film is water-soluble, it may easily be washed off and the re-hydration of the specimen is a very simplev operation.
- the cellulose ether composition is stable and is easy to use. Certain of the specimens smeared onto the slide may be allowed to dry in the air, others may be dehydrated, but the re-hydration makes certain that the specimens are in good condition for examination,
- cellulose i ether There may be used in the solution of the cellulose i ether different proportions of solvents and in many cases only a single solvent may be used and the concentration of the cellulose ether may be varied. In place of the cellulose ether, other materials having similar'properties may be used or one may apply two substances, one of which is film-forming and the other of which has surface active properties. In re-hydrating the cells, in some instances only water need be used but in order to facilitate the operation and facilitate the examination, the surface acting agent should be present as in such case the re-hydration requires not over about a minute.
- a method of treating slides having thereon smears of body cells for microscopic examination which comprises providing a solution of a Water soluble synthetic film forming material in an ether-alcohol solvent, dropping said solution on said slide to fix said cells, allowing said solvent to evaporate, thereafter treating said slide with an aqueous liquid to dissolve said material, and staining said slide preparatory to microscopic examination.
- a method of treating slides having thereon smears of body cells for microscopic examination which comprises providing a solution of a water soluble synthetic film forming material in an ether-alcohol solvent, allowing said smears on said slide to dry and then dropping said solution on said slide, allowing said solvent to evaporate, thereafter treating said slide with an aqueous liquid to dissolve said material, and staining said slide preparatory to microscopic examination.
- a method of treating slides having thereon smears of body cells for microscopic examination which comprises providing a solution of a water soluble synthetic film forming material in an ether-alcohol solvent, dropping said solution on said slide, said material being an alkyl cellulose ether, allowing said solvent to evaporate, thereafter treating said slide with an aqueous liquid to dissolve said material and to rehydrate said cells, and staining said slide preparatory to microscopic examination.
- a method of treating slides having thereon smears of body cells for microscopic examination which comprises providing a solution of a water soluble synthetic film forming material in an ether-alcohol solvent, said material being an alkyl cellulose ether, dropping said solution on said slide to dehydrate said cells, allowing said solvent to evaporate, thereafter treating said slide with an aqueous liquid to dissolve said material and to re-hydrate said cells, and staining said slide preparatory to microscopic examination.
- a method of treating slides having thereon smears of body cells for microscopic examination which comprises providing a solution of a water soluble synthetic film forming material in an ether-alcohol solvent, said material being hydroxy propyl methyl cellulose ether, dropping said solution on said slide to dehydrate said cells, allowing said solvent to evaporate, thereafter treating said slide with an aqueous liquid to dissolve said material and to re-hydrate said cells, and staining said slide preparatory to microscopic examination.
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Physics & Mathematics (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Sampling And Sample Adjustment (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Description
United States Patent 9 TREATMENT OF MICROSCOPE SPECIMEN SLIDES Herbert E. Nieburgs, New York, N.Y.
No Drawing. Application June 3, 1957 Serial No. 663,007
8 Claims. (Cl. 167-845) The present invention is directed to the subject of microscopic examination of body cells and more particularly to a method of treating the transparent slides in order to preserve the specimens for such examination.
It has, of course, been common practice to place a smear on a slide and examine it under the microscope. If such smear is allowed to dry in the air, the cells under certain conditions are not suitable for obtaining a clear picture under the microscope. To avoid this the specimen smear is fixed in a mixture of ether and alcohol. However, this is not always done soon enough so that some dehydration takes place. If the slide is to be transferred to the laboratory by mail or other means, the surface of the smear was usually protected by glycerine. However, this could very easily be rubbed off in whole or in part in packing, handling or during shipment and the specimen may thus be damaged. There is also the danger of detachment of cells during the removal of the glycerine in the laboratory.
In order to overcome these deficiencies and disadvantages, it is among the objects of the present invention to provide a method of protecting and fixing the specimen on the slide simultaneously in a single step whereby the slide may be handled without danger of damage to the specimen.
It is also among the objects of the present invention to provide a method wherein a dried and non-fixed specimen may be rehydrated at the point of use, thus affording the analyst a clear picture of the chromatin.
It is further among the objects of the present invention to provide a composition for carrying out the method, which composition is stable, is easily handled and is highly effective in its application to the slide.
In practicing the present invention there is provided a solution of a water-soluble synthetic film-forming material which preferably also has the property of reducing surface tension. The solvent used is generally a mixture of ethyl ether and ethyl alcohol.
The specimen smear on the slide is treated with the solution by flowing said solution onto the slide to cover the specimen. It is important that the slide be not dipped into the solution as otherwise at least part of the cells of the specimen may be washed off. The solvent is allowed to evaporate, thereby not only fixing the cells but also forming a film thereon to protect it during transit. Before examining the specimen, the slide is then treated with an aqueous ether alcohol liquid which dissolves the film.
Thereafter the slide is stained as usual and is subjected to examination under the microscope.
It has been found that an alkyl cellulose ether soluble in water is an ideal substance for the purpose. Specifically there is provided for immediate fixation and surface coating a capsule containing .2 gram of hydroxy propyl methyl cellulose ether, and dissolving the cellulose ether in 10 cc. of water at a temperature of, say, 80 to 90 C. To this solution is added 190 cc. of a mixture of in 3 parts of water to 2 parts of a mixture of equal equal parts of ethyl alcohol and ethyl ether. The cellulose ether preferably has a viscosity of about 4,000 centipoises although cellulose ethers of other viscosities are also usable.
While in the above description the cellulose ether is in powder form Within a capsule, it is highly feasible to provide a complete solution as stated above in a suitable container such as an ampule, jar, bottle or the like for convenience of the analyst. In such case the analysts preparation of the slide is minimized.
In taking the specimen for examination, a cervical or a vaginal material is placed upon the slide. The specimen may be allowed to dry in the air. In the laboratory the slide is treated with the use of a solution of a surface active agent in water in a suitable concentration. It has been found that the sorbitol or mannitol partial esters of higher fatty acids such as those having 16 and 18 carbon atoms are suitable for the purpose. The specimen is then fixed, stained and examined microscopically.
For rehydration of dried unfixed specimens the capsule containing .2 g. hydroxy propyl methyl cellulose ether has the property of reducing the surface tension of water to about 43 dynes per centimeter, in a solution of 400 cc. of water and 200 cc. of a mixture of 95% ethyl alcohol and ethyl ether. Dried and unfixed slides are dipped in this solution for about five minutes, fixed in ether-alcohol for about ten minutes and then stained.
In the case of endometrial material, or that of gastric origin, breast, urine and others, a somewhat different procedure may be used. The specimen on the slide as soon as the smear is made, is fixed by a mixture of ethyl ether and ethyl alcohol while still fresh usually by dropping said solution upon the specimen on the slide. It is allowed to dry whereby fixation of the cells is obtained and a film formed thereon to protect it. This immediate fixing is important for such specimens since destruction of cells occurs on standing even for a relatively short time. The fixed specimen may be taken to the laboratory for examination. There the slide is placed amounts of ether and alcohol to remove the protective film, stained as usual and examined microscopically.
Although the invention has been described setting forth a single cellulose ether composition, it is apparent that other substances having the desired properties may be used in place thereof. When the specimen has been treated with the solution of .2 g. of cellulose ether in 10 cc. of water and cc. of a mixture of ether and alcohol, it requires, say, twenty to thirty minutes for evaporation of the solvent and it leaves a thin hard film over the specimen. The cells have been fixed and the film protects the material during handling and transit. Therefore, the specimen is in the optimum condition for the microscopic examination. Gastric specimens and those obtained from duodenal drainage undergo rapid enzymatic digestion. If placed in ether-alcohol for fixation while wet, detachment of cells may occur, which is eliminated by this method.
There are a number of advantages inherent in the present invention in that the flowing of the cellulose ether solution onto the slide prevents the washing away of cells and after drying the specimen may be handled even carelessly without damage to the specimen. Because the film is water-soluble, it may easily be washed off and the re-hydration of the specimen is a very simplev operation. The cellulose ether composition is stable and is easy to use. Certain of the specimens smeared onto the slide may be allowed to dry in the air, others may be dehydrated, but the re-hydration makes certain that the specimens are in good condition for examination,
There may be used in the solution of the cellulose i ether different proportions of solvents and in many cases only a single solvent may be used and the concentration of the cellulose ether may be varied. In place of the cellulose ether, other materials having similar'properties may be used or one may apply two substances, one of which is film-forming and the other of which has surface active properties. In re-hydrating the cells, in some instances only water need be used but in order to facilitate the operation and facilitate the examination, the surface acting agent should be present as in such case the re-hydration requires not over about a minute.
While the invention has been described setting forthseveral specific embodiments thereof, the invention is not limited thereto as various changes. in the details may be made within the spirit of the invention and such variations are within the skill of the ordinary worker in the field. Therefore, the invention is to be broadly construed and not to be limited except by the character of claims appended hereto.
I claim:
1. A method of treating slides having thereon smears of body cells for microscopic examination which comprises providing a solution of a Water soluble synthetic film forming material in an ether-alcohol solvent, dropping said solution on said slide to fix said cells, allowing said solvent to evaporate, thereafter treating said slide with an aqueous liquid to dissolve said material, and staining said slide preparatory to microscopic examination.
2. A method according to claim 1 in which said synthetic material is hydroxy propyl methyl cellulose ether.
3. A method of treating slides having thereon smears of body cells for microscopic examination which comprises providing a solution of a water soluble synthetic film forming material in an ether-alcohol solvent, allowing said smears on said slide to dry and then dropping said solution on said slide, allowing said solvent to evaporate, thereafter treating said slide with an aqueous liquid to dissolve said material, and staining said slide preparatory to microscopic examination.
4. A method according to claim 3 in which said'synthetic material is hydroxy propyl methyl cellulose ether.
5. A method of treating slides having thereon smears of body cells for microscopic examination which comprises providing a solution of a water soluble synthetic film forming material in an ether-alcohol solvent, dropping said solution on said slide, said material being an alkyl cellulose ether, allowing said solvent to evaporate, thereafter treating said slide with an aqueous liquid to dissolve said material and to rehydrate said cells, and staining said slide preparatory to microscopic examination.
6. A method of treating slides having thereon smearsi hydroxy propyl methyl cellulose ether, allowing said solvent to evaporate, thereafter treating said slide with an aqueous liquid to dissolve said material, and staining said slide preparatory to microscopic examination.
7. A method of treating slides having thereon smears of body cells for microscopic examination which comprises providing a solution of a water soluble synthetic film forming material in an ether-alcohol solvent, said material being an alkyl cellulose ether, dropping said solution on said slide to dehydrate said cells, allowing said solvent to evaporate, thereafter treating said slide with an aqueous liquid to dissolve said material and to re-hydrate said cells, and staining said slide preparatory to microscopic examination.
8. A method of treating slides having thereon smears of body cells for microscopic examination which comprises providing a solution of a water soluble synthetic film forming material in an ether-alcohol solvent, said material being hydroxy propyl methyl cellulose ether, dropping said solution on said slide to dehydrate said cells, allowing said solvent to evaporate, thereafter treating said slide with an aqueous liquid to dissolve said material and to re-hydrate said cells, and staining said slide preparatory to microscopic examination.
References Cited in the file of this patent UNITED STATES PATENTS 2,265,913 I Lillienfeld Dec. 9, 1941 2,602,755 Silvernail July 8, 1952 2,648,165 Nestor Aug. 11, 1953
Claims (1)
1. A METHOD OF TREATING SIDES HAVING THEREON SMEARS OF BODY CELLS FOR MICROSCOPIC EXAMINATION WHICH COMPRISES PROVIDING A SOLUTION OF A WATER SOLUBLE SYNTHETIC FILM FORMING MATERIAL IN AN ETHER-ALCOHOL SOLVENT, DROPPING SAID SOLUTION ON SAID SLIDE TO FIX SAID CELLS, ALLOWING SAID SOLVENT TO EVAPORATE, THEREAFTER TREATING SAID SLIDE WITH AN AQUEOUS LIQUID TO DISSOLVE SAID MATERIAL, AND STAINING SAID SLIDE PREPARATORY TO MICROSCOPIC EXAMINATION.
Publications (1)
| Publication Number | Publication Date |
|---|---|
| US2899363A true US2899363A (en) | 1959-08-11 |
Family
ID=3448164
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| US2899363D Expired - Lifetime US2899363A (en) | Treatment of microscope specimen slides |
Country Status (1)
| Country | Link |
|---|---|
| US (1) | US2899363A (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US3493447A (en) * | 1963-12-04 | 1970-02-03 | Peter Adrian Rock | Method of and apparatus for preparation of specimens for microscopic examination |
| US4001460A (en) * | 1974-04-22 | 1977-01-04 | Kinney Thomas D | Light microscopy processing method |
| FR2320543A1 (en) * | 1975-08-08 | 1977-03-04 | Gen Electric | PROCESS FOR COLORING AND SEALING A BIOLOGICAL SAMPLE |
| FR2320544A1 (en) * | 1975-08-08 | 1977-03-04 | Gen Electric | FORMULATION AND PROCESS FOR COLORING AND SEALING A BIOLOGICAL SAMPLE |
| US4221823A (en) * | 1978-04-21 | 1980-09-09 | Phillips Petroleum Company | Process for mounting palynological specimens |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US2265913A (en) * | 1933-07-17 | 1941-12-09 | Lilienfeld Patents Inc | Hydroxy alkyl-alkyl ethers of cellulose |
| US2602755A (en) * | 1951-01-02 | 1952-07-08 | Dow Chemical Co | Thermoplastic compositions of water-soluble cellulose ethers |
| US2648165A (en) * | 1945-06-04 | 1953-08-11 | Minnesota Mining & Mfg | Seed carrier |
-
0
- US US2899363D patent/US2899363A/en not_active Expired - Lifetime
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US2265913A (en) * | 1933-07-17 | 1941-12-09 | Lilienfeld Patents Inc | Hydroxy alkyl-alkyl ethers of cellulose |
| US2648165A (en) * | 1945-06-04 | 1953-08-11 | Minnesota Mining & Mfg | Seed carrier |
| US2602755A (en) * | 1951-01-02 | 1952-07-08 | Dow Chemical Co | Thermoplastic compositions of water-soluble cellulose ethers |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US3493447A (en) * | 1963-12-04 | 1970-02-03 | Peter Adrian Rock | Method of and apparatus for preparation of specimens for microscopic examination |
| US4001460A (en) * | 1974-04-22 | 1977-01-04 | Kinney Thomas D | Light microscopy processing method |
| FR2320543A1 (en) * | 1975-08-08 | 1977-03-04 | Gen Electric | PROCESS FOR COLORING AND SEALING A BIOLOGICAL SAMPLE |
| FR2320544A1 (en) * | 1975-08-08 | 1977-03-04 | Gen Electric | FORMULATION AND PROCESS FOR COLORING AND SEALING A BIOLOGICAL SAMPLE |
| US4221823A (en) * | 1978-04-21 | 1980-09-09 | Phillips Petroleum Company | Process for mounting palynological specimens |
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