By Carl C Lindegren
Sm 4to, 1949, ,
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E. Distilled water, rinse. f. 50% alcohol, 2 minutes. g. Fast green in 95% alcohol (enough fast green to color the alcohol faintly), 3-5 minutes. - Absolute alcohol, rinse. Clear, two 2-5 minute changes in toluene, mount. Results: Chromosomes - red Centrosome - red Nucleolus - pale green Cytoplasm - pale green 5. Crystal violet-anilin blue. This stain combination used after the I and KI mordant proved an excellent one for showing the paired h. i. chromosomes. a. Fixation, 8-10 minutes. b. Tap water wash, 10-15 minutes.
C. d. e. f. g. Fixation, 8-10 minutes. Tap water wash, 10 minutes. 5% solution of sodium thiosulfate, 10 minutes. 3-5 minutes. water, Distilled Stain in 1-20 Giemsa -distilled H2O solution, 15-40 minutes. Distilled water, rinse. Dehydrate and clear in following acetone -toluene steps, checking progress with microscope: (a) pure acetone 95 parts, toluene 5 parts; (b) acetone 70 parts, toluene 30 parts; (c) acetone 30 parts, toluene 70 parts, (d) pure toluene; then mount in balsam. Wash STAINING TECHNIQUES Results: Chromosomes Centrosome Nucleolus 8.
Rafalko (1945) devised a modified Feulgen stain with which the yeast chromosomes were successfully stained for the first time. The procedure Is as follows: The leuco basic fuchsln Is prepared by gentle bubbling of sulphur dioxide gas from a small aperture In glass tubing Into 1/2% solution of basic fuchsln until decolorlzatlon occurs (In about 10-30 minutes). The solution Is then ready for use. Distilled water Is similarly saturated for the sulphurous acid rinsing bath and may be stored for many weeks In a tightly -corked flask.