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KMID : 0378019610040110037
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New Medical Journal
1961 Volume.4 No. 11 p.37 ~ p.40
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On the Fat-staining of Peripheral Human Blood Cells.
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Abstract
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1) Fixation.
Lay 8-10 folds of gauze in a Petri´s dish and put two glass rods parallelly on them. Drop formol or formolalcohol solution (neutral formol stock solution three parts, 95% ethylalcohol one part), and let the formol or formol-alcohol to evaporate and fill up the capped Petri´s dish. Make thin smear and lay it on the glass rods immediately after the smear is madewhile still wet. The smeared surface should be faced downwards. Fix it for 10-20 seconds.
Presence of clouding due to evaporated water in the cover indicates that the fixation is not satisfactory. The gauze may be used repeatedly after careful drying.
2) Allow the smear to dry in air for 30-60 minutes at room temperature,
3) Dilute stock dye solution two times wifl ´equal amount of distilled water, pour it on She smear:soon.. after filtration, and stain it for 3-5 minutes at room temperature.
4) Rinse away with three time diluted -95% ethylalcohol solution until the dye granules on the surface of the smear are unvisible with the naked eye.
5) Wash the smear with tap water.
6) Stain the nucleus with Boemer´s hematoxilin for 5-10 minutes.
7) Keep it in running water for about 10 minutes.
8) Seal it up by levulose-syrup or fluid honey.
The stock dye solution
After dissolving 0.5 gm of naphtholum alpha- medicinale recryst. (merck) completely with 20 cc of 95% ethylalcohol in a clean -tube, add 0.5 gm of ground sudan III (Gruebler). Closing tightly, shake the glass tube strongly to mix the dye. Place the mixure overnight at room temperature. Dilute with 5cc of distilled water next day and store the solution at cool place. The stock dye solution can be used for about 10 days from 5-7 days after it is made.
The selection of dye and naphthol is necessary; Sudan III (Gruebler) is the best among sudan dyes. When we cannot get this dye there is nothing for it but to use scharlach Rot(Merck) or sudan IV(Coleman & Bell). Oil red O(national aniline division),. Sudan Schwarz B(Merck), Sudan III (Merck). etc. are not useful. Naphtholum alpha -medicinale recryst. (Merck) is the best. Beta-naphthol is not useful.
Result
All the eosinophilic granulocytes and myelocytes are full of fat-granules. The granules are large and red in color, whose center is slightly paler than the periphery.
All the neurtophilic granulocytes and myelocytes are also full of fat-granules. The granules are smaller than those of cosinophils and dark-red in color.
The basophils of normal blood show fat-granules in 20%. The basophils and basophilic.myelocytea: sess two kind of granulesthe positive and the negative, The former is dark-red in color, the transparent.
All the monocytes reveal fat-granules. However, the granules are usually less in number than in neutron phils and distribuled relatively uniformly in the majority of the cells.
The majority of myeloblasts possess fat-granules whose number is variable in each cell.
Most of the paramyeloblasts also show fat-granules. The granules in them are usually less in number as in monocytes, but the polymorphonuclear cells reveal only a few or even no gat-granules. . The Auer´s bodies are also dark-red in color, whose center is slightly paler than the periphery.
The reticulum cells exhibit small fat-granules; most of the lymphoid cells, the spindle forms, and the starry cells show no fat-granules, but all the monocytoid cells possess fat-granules which tend to be arranged around the nucleus.
The cells of lymphatic series also reveal fat-granules; The lymphocytes of normal blood are positive in 80%, but the positive cells decrease by far in lymphatic leukemias.
All the thrombocytes also possess small fat-granules.
All the erythrocytes as well as erythroblasts show no fatgranules.
The hematokonies areafat-stainable and round or over in shape. They are relatively rarely found in the plasm of normal blood. On the other hand, they appear in great number in chronic myeloid leukemias, but they are not found in lymphatic leukemias and leukemic reticuloendoteliosis.
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KEYWORD
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