BENSLEY'S NEUTRAL 



39 



BEST'S CARMINE 



Bensley's Neutral Safranin. For mitochon- 

 dria and secretion antecedents especially 

 in the pancreas. Fix in 2.5% aq. potas- 

 sium bichromate, 100 cc. ; mercuric 

 chloride, 5 gms. 24 hrs. Wash, dehy- 

 drate, clear, imbed and section. To 

 prepare stain slowly add sat. aq. acid 

 violet to sat. aq. safranin O in a flask 

 until ppt. ceases when a drop of mixture 

 on filter paper gives not an outside red 

 rim of safranin but a solid neutral color. 

 Filter. The filtrate should be as nearly 

 as possible colorless. Dry ppt. on filter 

 paper and make of it a sat. sol. in abso- 

 lute alcohol. Pass sections through 2 

 changes each of toluol and absolute 

 alcohol, then down through lower alco- 

 hols to aq. dest. (Bleach chrome and 

 osmium fixed tissues in permanganate 

 and oxalic acid, as described under 

 Anilin Fuchsin Methyl Green and 

 mercury fixed ones in Lugol's solution, 

 10 sec. finally washing in aq. dest.) 

 Dilute alcoholic neutral safranin with 

 equal volume aq. dest. and stain 5 min.- 

 2 hrs. Quickly blot with filter paper. 

 Plunge into acetone and immediately 

 pass to toluol without draining. Exa- 

 mine and if further differentiation is 

 needed treat with oil of cloves. If this 

 is not enough rinse in abs. ale, flood 

 momentarily with 95% ale. and pass 

 back through absolute to toluol. Wash 

 in 2 changes toluol and mount in balsam. 

 This is a difficult method but the results 

 are worth it. (see Bensley, R. R., 

 Am. J. Anat., 1911, 12, 297-3S8). 



Benzamine Blue 3B, see Trypan Blue. 



Benzene-Azo-a-Naphthylamine. A mono- 

 azo dye used bv Carter, J. S., J. Exp. 

 Zool., 1933, 65/159-179 as a vital stain 

 for Stenostomum. 



Benzo Blue 3B, see Trypan Blue. 



Benzo New Blue 2B, see Dianil Blue 2R. 



Benzo Sky Blue, see Niagara Blue 4B. 



Benzoazurine G (CI, 502), a direct dis-azo 

 dye of light fastness 4 sometimes 

 polychromatic (nuclei red, cytoplasm 

 blue to blue-violet). Applied after 

 treating blue-green algae with copper 

 sulphate, spores orange red, vegetative 

 cells dark blue or violet (Emig, p. 41). 



Benzoin Blue R, see Azo Blue. 



Benzopurpurin 4B (CI, 448) — cotton red, 

 diamin red, dianil red, Sultan and 

 direct red, all 4B — An acid dis-azo dye 

 no longer used. 



Benzyl Benzoate is employed in the Spalte- 

 holz Method of clearing. 



Benzyl Violet. Conn (p. 132) states that 

 this term relates to a group of violets 

 which are pararosanilins, some acid and 

 some basic, with benzyl substitution in 

 one or more amino groups. 



Berberian's Method. Berberiau, D. A., 

 Arch. Dermat. & Syphil., 1937, 36, 1171- 



1175, has (iv-^veloped a method for stain- 

 ing fun;.';i in epidermal scales and hair, 

 which differentiates epithelial cells, 

 blood cells, bacteria and 'mosaic 

 fungi'. The following account was 

 written by D. A. Berberian, American 

 University of Beirut, Beirut, Lebanon, 

 June 22, 1946: 



Fix small pieces of scales or hair on a 

 slide with 50% aq. glacial acetic acid 

 by drying in an incubator. Defat, 

 clear, hydrate, and wash off the acid as 

 follows: Cover the preparation with 

 ether 2-3 times, 20-30 sec. each; flood 

 twice with absolute acetone, 30-60 sec. 

 each; and then flood consecutively with 

 absolute, 95, 70 and 50 per cent alcohol. 

 Stain for 3-5 min. with Marti notti's 

 solution (aq. dest., 75 cc; lithium car- 

 bonate, 0.5 gm.; and toluidine blue, 

 1 gm. After the stain dissolves, add 

 20 cc. glycerin and 5 cc. 95% alcohol). 

 Wash gently in water and differentiate 

 with 0.5% acetic acid. Dehydration is 

 best carried out by 3-4 changes of 

 absolute acetone kept 2-3 minutes each 

 time. Pass through xylol and mount 

 in Euparol or any other neutral mount- 

 ing agent. Success of preparation 

 depends largely on proper differentia- 

 tion, dehydration and de-acidification. 

 See Fungi. 



Berberine Sulphate. x\n alkaloid used as a 

 fluorochrome for malarial parasites 

 (Metcalf, R. L. and Patton, R. L., 

 Stain Tecbn., 1944, 19, 11-22). 



Bergamot Oil is sometimes used for clearing 

 because it wall mix with 95% alcohol. 



Berlin Blue is another name for Prussian 

 Blue (a metallic pigment). It is em- 

 ployed for microchemical detection of 

 Iron. Kremer, Zeit. f. wiss. mikr., 

 1938, 54, 429-432 suggests proceeding as 

 follows: Fix in absolute alcohol. De- 

 paraffinize lOp sections. Bleach in 3-5% 

 H2O2 3-5 days. Wash carefully in aq. 

 dest. Quickly darken in (NH4)2S. 

 Transfer to K ferrocyanide and HCl. 

 Iron gives blue color. 



Curiously enough when injection of 

 blood vessels is demanded this mineral 

 pigment is usually called for as Berlin 

 blue. Thus the Bensleys (p. 153) give 

 directions for making up Tandler's 

 Berlin blue gelatin. Soak and melt 5 

 gms. pure gelatin in 100 cc. aq. dest. 

 Add sufficient Berlin blue to give desired 

 color and then 5-6 gms. potassium iodide 

 and a crystal of thymol as a preservative. 

 Inject this mass, which is fluid above 

 17 °C. Fix tissues in 5% formalin which 

 preserves it even through decalcifica- 

 tion. 



Best's Carmine. Griibler's oarminum ru- 

 brum optimum, or some other good 

 carmine, 2 gm.; potassium carbonate, 



