416 methods and formulas ds 21.3 



21.3 Special Stains for Blood 



Nowhere is the purpose of the present writer in his classification of staining tech- 

 niques more likely to be misunderstood than in tliis division. At least a hundred methods 

 for staining blood liave been advocated, and it is today conventional to utilize only the 

 methods given in Chapter 20 (DS 13.1, 13.2, and to a less extent 13.3). Those techniques 

 are, however, of far wider application, for they may be used for sections, for staining 

 parasites in blood, and for many other purposes. Moreover, blood films themselves may 

 be excellently stained by almost any triple-staining method, though it usually is a 

 surprise to the hematologist who has seen nothing but a methylene blue-eosin prepara- 

 tion to observe the magnificent differentiation which can be secured by many of the 

 DS 12.3 or DS 13.4 techniques. Under these circumstances the writer has included in the 

 present section only those methods of staining blood which are useless for any other 

 purpose. 



21.3 Brice 1930 see DS 22.8 Brice 1930 



21.3 Bruner and Edwards 1940 see DS 23.219 Bruner and Edwards 1940 



21.3 Bacsich 1936 11025, 70:267 



REAGENTS REQUIRED: A. water 20, 95% ale. 80, phenol 2.5, Sudan III 0.5; B. DS 11.121 



Weigert 1903; C. 0.5% hydrochloric acid in 40% ale. 

 PREPARATION OF A: Boil the dye 5 minutes in 100 95% ale. Add phenol and filter hot. 



Cool to 4°C., 24 hours. Filter. Add water drop by drop till ale. eoneentration reduced 



to 80%. 

 method: [air-dried smears] — > A, on slide, 5 mins., 56°C. -^ thorough wash — > B, 30-00 



sees. -^ rinse — > C, till differentiated -^ wash — * dry -* M 12.1 Zwemer 1933 



21.3 Baillif and Kimbrough 1947 11284, 32:155 



REAGENTS REQUIRED: A. sat. sol. Sudan black B in 70% ale.; B. DS 13.11 May and 



Grunwald 1902; C. DS 13.13 Giemsa 1902 2, water 98 

 method: [ale. -formaldehyde-fixed smears]—* A, 30-60 mins.-* rinse—* B, on slide, 3 



mins. — * add water, 1 min. — * drain -^ C, on slide, 15 mins. -^ wash — > dry 



21.3 Buzard 1930 4285a, 7 :264 



REAGENTS REQUIRED: A. 1% acid fuchsin; B. 0.8% bromine water; C. 1% hydrochloric 



acid in 95% ale. 

 methods: [fixed smears] — > A, 30 sees. — > B, 4-5 mins. -^ wash -^ C, till mauve -^ bal- 

 sam, via usual reagents 



21.3 Campbell and Alexander test. 1938 Mallory Mallory 1938, 257 



REAGENTS REQUIRED: A. acctic acid 0.5, benzidine 0.1, sodium nitroferrieyanide 0.1; B. 

 water 100, 30% hydrogen peroxide 0.1 



PREPARATION OF A: Dissolvc benzidine in acetic acid and dilute to 20. Dissolve nitro- 

 ferrieyanide in 20 water and add to benzidine. Dilute mixture to 100. Filter. 



method: [200 to 300 m frozen sections of formaldehyde-fixed material]^ A, 30 mins. 

 37°C. with frequent shaking — * rinse -^ B, 30 mins., 37°C. with frequent shaking — > 

 balsam, via usual reagents 



recommended for: capillaries in brain. 



result: capillaries black against colorless background. 



21.3 Crossmon 1940 20540b, 15:155 



REAGENTS REQUIRED: A. water 100, acetic acid 1, chromotrope 2R 0.25; B. 5% phospho- 

 tungstic acid in 95% ale.; C. water 100, aeetie acid 1, methyl blue 0.5; D. 2% acetic 

 acid 

 method: [sections from formaldehyde-fixed material] -^ water — » A, 1-5 mins. -^ rinse 

 -^ B, till erythrocytes alone stained — » C, 2-5 mins. -^ D, till differentiated — > bal- 

 sam, via usual reagents 

 recommended for: differential staining of erythrocytes in sections. 



