• 专利附录
  • 专利说明
  • 权利要求
  • 类似技术
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  • 引用文献
  • 法律状态
  • 优先权
    • 专利摘要:
    1480576 Blood stain composition BOEHRINGER MANNHEIM GmbH 6 April 1976 [11 April 1975] 13853/76 Heading G1B [Also in Divisions C4 and G2] In blood investigation superior results are obtained using a pre-coated slide having thereon a mixture of methylene blue N and cresyl violet acetate if (a) they are chromatographically pure and (b) if used in the ratio 1 : 1À5 to 1 : 5. The methylene blue N is preferably used in the form of its monohydrochloride. The purification is described, [see Group C4].
    公开号:SU904536A3
    申请号:SU762346753
    申请日:1976-04-08
    公开日:1982-02-07
    发明作者:Бергер Дитер；Гютляйн Вернер；Вернер ВОЛЬФГАНГ；Рикманн Петер
    申请人:Берингер Маннхайм Гмбх (Фирма)；
    IPC主号:
    专利说明:

    (5) SUBJECT GLASS FOR RESEARCH The invention relates to medicine, namely to hematology. A glass slide is known for the following and blood with a dye layer deposited on it consisting of methylene blue and sodium acetate in a ratio of 9: 1 tn. However, dyes applied to a glass slide do not provide high contrast staining (dim color), and the forms of granular leukocytes (eosinophils, neutrophils, and basophils) differ with great difficulty. The purpose of the invention is to increase the coloring contrast. This goal is achieved by the fact that in a glass slide for examining blood coated with a layer of dyes of methylene blue and acetate, the cresolviolet layer of dye contains dyes in the cooler wearing 1: 1.5 - 1: 5. In this case, methylene blue is used as monohydrochloride. Methylene solution, 1L is purified by dissolving in water, the hydrochloride is separated with hydrochloric acid. A chromatographically pure product is obtained which contains only about 0.5 zinc chloride and is presented as mono-hydrochloride. The cresolviolet acetate is purified by carefully settling in water, removing the sodium acetate, then dissolved in methanol and isolated with ether. Repeating this process gives a chromatographically pure product. The dyes thus purified are dissolved in B methanol and applied to a glass slide. They can also be dissolved in water and applied along with polyoxyethylene sorbitan monopalmitate. Ratio of these coloring substances 1: 1.5 1: 5390 The dye is applied on a glass slide from 0.5 to. A layer of dye is then applied to the coated glass, a drop of blood is added, covered with a cover glass, and after 3-5 minutes it is microscoped with an immersion objective. A bright contrasting color is obtained, the reproduction is good. Eosinophil granules are colored bright yellow, basophil granules orange. The contours of the nuclei are clearly recognized, which greatly facilitates the differentiation of lymphocytes, monocytes and various mature stages of granulocytes. Example 1. Purification of cresolviolet acetate. 50 g of cresolviolet acetate are suspended in 100 ml of water and mixed vigorously for 15 min. Then the edges of the substance are sucked off and washed twice with 100 ml of ice-cold water. The filtered precipitate is dissolved in 1.6 L of methanol with heating and sucked off from insoluble particles at 30 ° C. The filtrate is slowly mixed with 3.5 L of ether and then stirred for 30 minutes while cooling on ice. The resulting crystals are sucked off, dissolved when heated in 620 ml of methanol, then the solution is cooled to 30 ° C and the dye is isolated by adding 1.85 liters of ether while stirring and cooling on ice. After suction and three-phase washing with 120 ml of ether-methanol C3.1), after drying, over diphosphoropentoxide, g of cresolviolet acetate in a photo of dark green crystals is obtained. Ingredients: about 821 cresol violet acetate, 8 cresol violet chloride and 10 water. When heated to 150 ° C, decomposition occurs. The substance according to chromatographic examination data is practically pure. The value of Rl 0.6. Example 2 “Purification of methylene blue. In a four-liter three-neck flask, 100 g of methylene blue in 1 l of water is dissolved with stirring and stirred with 1 l of concentrated. hydrochloric acid, 1.18. After four hours of stirring while cooling on ice and staying eight hours in the refrigerator at + 5 ° C, the resulting crystals are sucked off, washed with 00 ml of ice-cold 6N. hydrochloric acid and 500 ml of ether. 36.5 g of black crystals with a metallic luster are obtained. The purified dye consisted. it is from methylene blue hydrochloride chloride and contains 3K water. There are only traces of zinc. ka The dye according to the chromatographic examination is practically pure. The value of Rf 0.5. Example 3. Using dyes purified according to examples 1 and 2, prepare a solution for the following compounds: Methylene blue hydrochloride, mg 130 cresolviolet acetate, mg270 Methanol, ml (Ecolo 100 This solution with a cotton swab is applied or sprayed on a glass slide and dried. With an average droplet of about 20 H, the applied amount of the coloring matter is about 3. A drop of blood is applied to the glass slide approximately 5 10 jM and covered with a cover glass. After a min, the color is examined under a microscope at 800-fold magnification using an immersion lens. Separate blood particles are presented in the following coloration. Reticulocytes: a purple 4th mesh within barely painted red blood cells Neutrophils: a purple core inside a fine-grained almost colorless plasma. magenta inside the average size of a compact-granular orange-red plasma. Lymphocytes: a draw of purple color with a light magenta plasma. Monocytes: as lymphocytes, only larger and more plasma.
    Platelets: small purple particles.
    The proposed dye allows to increase the contrast of staining in the same way, when comparing stained slides at a ratio of 1: 1.5 to 1: 3 and 1: 5 with stained slides with a ratio of dyes
    9: 1 the color of the individual blood cells is different.
     The table shows the color of individual blood elements, depending on the ratio of methyl blue N-hydrochloride: cresyl violet acetate.
    As can be seen from the table, the use of slides containing methylene blue and cresolviolet or methylene blue acetate in the form of minohydrochloride in a ratio of 1: 1.5 -5 allows to obtain better staining of various blood elements compared to the known dye in a ratio of 9: 1
    权利要求:
    Claims (1)
    [1]
    1. A slide for examining blood with a dye deposited on it consisting of methylene blue and cresolviolet acetate, because in order to increase the contrast of staining, the dye layer contains components in the following ratio 1: 1.5 1: 5.
    2, Glass according to claim 1, characterized in that methylene blue is used as monohydrochloro
    Sources of information taken into account in the examination
    1. US Patent No. 379659, cl. 117-12, 1972.
    类似技术:
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    同族专利:
    公开号 | 公开日
    ES446836A1|1977-06-01|
    DE2515966A1|1976-10-21|
    SE7602728L|1976-10-12|
    FR2307271B1|1980-03-28|
    AU1267776A|1977-10-13|
    GB1480576A|1977-07-20|
    US4070495A|1978-01-24|
    IT1059617B|1982-06-21|
    AT350190B|1979-05-10|
    CS205019B2|1981-04-30|
    DK145318C|1983-04-05|
    FR2307271A1|1976-11-05|
    BE840454A|1976-10-07|
    SE420355B|1981-09-28|
    DD123628A5|1977-01-05|
    DK145318B|1982-10-25|
    LU74738A1|1976-11-11|
    PL107576B1|1980-02-29|
    FI760911A|1976-10-12|
    CA1052675A|1979-04-17|
    DK149876A|1976-10-12|
    ATA262076A|1978-10-15|
    CH621629A5|1981-02-13|
    IE42552B1|1980-08-27|
    JPS51126893A|1976-11-05|
    DE2515966C3|1979-11-22|
    DE2515966B2|1979-03-22|
    HU176928B|1981-06-28|
    IE42552L|1976-10-11|
    AR210757A1|1977-09-15|
    FI59677B|1981-05-29|
    FI59677C|1981-09-10|
    NL7603628A|1976-10-13|
    引用文献:
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    GB1473945A|1973-05-24|1977-05-18|Gen Electric|Method for preparing slides for blood evaluation|
    US3906120A|1970-10-30|1975-09-16|Gen Electric|Method for preparing slides for blood evaluation|
    US3796594A|1970-10-30|1974-03-12|Gen Electric|Stain coated slides for differentially staining blood|
    IT966513B|1970-10-30|1974-02-20|Gen Electric|PRE-COLORED SLIDES FOR BLOOD TESTS|DE2651060C3|1976-11-09|1980-06-26|Boehringer Mannheim Gmbh, 6800 Mannheim|Differential diagnostic semen examination|
    US4193980A|1978-01-05|1980-03-18|Corning Glass Works|Dry preparation for reticulocyte staining|
    US4248821A|1979-07-25|1981-02-03|Dellen Adrian F Van|Method and device for embedding a specimen for microscopic examination|
    US4500509A|1981-03-11|1985-02-19|Lawrence Kass|Metachromatic dye sorption and fluorescent light emmisive means for differential determination of developmental stages of neutrophilic granulocytic cells and other leukocytes|
    US4400370A|1980-03-12|1983-08-23|Lawrence Kass|Metachromatic dye sorption means for differential determination of leukocytes|
    US4581223A|1980-03-12|1986-04-08|Lawrence Kass|Individual leukocyte determination by means of differential metachromatic dye sorption|
    CA1155041A|1980-04-21|1983-10-11|Michael E. Jolley|Fluorescent nucleic acid stains|
    US4606950A|1984-08-31|1986-08-19|Ellen M. Corbet|Method for assembling a floral arrangement|
    US5106744A|1990-11-01|1992-04-21|Cytocolor Inc.|Method of staining monocytes and compositions thereof|
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    法律状态:
    优先权:
    申请号 | 申请日 | 专利标题
    DE19752515966|DE2515966C3|1975-04-11|1975-04-11|Pre-stained slides for blood testing|
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