STAINS, ETC. 



1398 



STAINING REAGENTS 



mended for demonstrating the granules in Ehrlich's 

 Mastzellen. Tissues hardened in alcohol are stained 

 for several hours in a solution of dahlia- violet 2 gm., in 

 90 per cent, alcohol 25 c.c. , distilled water 70 c.c. , 

 glacial acetic acid 5 c.c. Wash in alcohol until near- 

 ly colorless. Deltapurpurin. See Benzopurpurin. 

 Dinitroso-resorcin. See Staining of Nerve-tissue, 

 Plainer' S Method. Eosin. Stains rapidly and gives 

 a beautiful, diffuse, rosy hue. It is one of the best con- 

 trast-stains with hematoxylin. Make a 5 per cent, aque- 

 ous solution, and dilute as required. It is also a spe- 

 cific stain for red blood-corpuscles (to which it gives a 

 copper color), for certain leukocytes, and for the giant- 

 cells of leprosy and tubercle. Erythrosin. See Eosin. 

 Fuchsin. Dissolve 1 gm. of fuchsin in 15 c.c. of dis- 

 tilled water, and 5° c - c - of 90 per cent, alcohol, and 

 add 100 c.c. of glycerin. This solution is used for 

 staining blood-corpuscles. Carbol- fuchsin : fuchsin 

 I gm. , carbolic acid 5 gm. , alcohol 10 gm. , distilled 

 water loo gm. Gentian-violet. A nuclear stain, 

 prepared by dissolving 0.5 gm. of the dye in 80 c.c. of 

 distilled water, 20 c.c. of 90 percent, alcohol, and gla- 

 cial acetic acid I c.c. Stains in 5 minutes. Dissolved 

 in indifferent media it may be used for staining intra 

 vitam, and in acid solutions colors the nuclei of fresh 

 tissues. It may be used according to Gram's method. 

 Hoffmann's Blue. A nuclear stain for sieve areas. 

 Dissolve I gm. of Hoffmann's blue in 20 c.c. of 90 per 

 cent, alcohol and 80 c.c. of distilled water, and add 0.5 

 c.c. of glacial acetic acid. Let sections remain in the 

 solution for about IO minutes. Indulin. See Nigro- 

 sin. Iodin-green. See Methyl-green. Magenta. 

 See Fuchsin. Manchester Brown. See Bismarck 

 Brown. Metanil-yellow. A plasmatic stain with a 

 special affinity for connective tissues. It is also used 

 to differentiate certain preparations ( Griesbach ). 

 Methylene-blue. An important reagent, which gives 

 a specific stain for lymph-spaces and intercellular ce- 

 ment, closely resembling gold and silver impregnation, 

 for medullated nerves, and for plasma-cells. It also 

 stains intra vitam, and is a specific reagent for the axis- 

 cylinders of sensory nerves in living animals (Ehrlich). 

 Small and permeable aquatic organisms may be stained 

 during life by adding to the water containing them 

 enough of the dye to give it a very pale tint. Nerve- 

 tissue may be stained by injecting the dye into the vas- 

 cular system of a living, narcotized animal, or by re- 

 moving the organ and immersing it in the solution. 

 From one-half to 1 per cent, solutions in physiologic salt- 

 solution are employed for this purpose. The color is 

 not permanent, but may be fixed by ammonium picrate. 

 Parker fixes the color by dehydrating in a solution of 

 mercuric chlorid, I gm. , in methylal 5 c.c. ; washing in 

 a mixture of 2 parts of the methylal and sublimate solu- 

 tion, I part pure methylal, 3 parts xylol. The object 

 is then placed in xylol for 4 or 5 days, when it is ready 

 to mount or embed. Mayer's albumen should not be 

 used to fix sections to the slide, as it discharges the 

 color. A solution of 0.25 gm. in 90 per cent, alcohol, 

 20 c.c. , and distilled water, 80 c.c, is used for tissue- 

 staining. One per cent, and saturated alcoholic (15 

 gms. to 100 c.c.) solutions are used for staining micro- 

 organisms. Dogiel's Methylene-blue and Ammo- 

 nium Picrate. Stain for 10 minutes in a 4 per cent, 

 solution of methylene-blue in normal salt-solution ; soak 

 for ]/ 2 an hour or more in a saturated solution of ammo- 

 nium picrate, wash in fresh ammonium picrate, and ex- 

 amine in dilute glycerin. This process is used to demon- 

 strate epithelia and lymph-spaces, and has the effect of 

 negative impregnation with silver nitrate. A positive 

 image is obtained by a short bath in the stain, a negative 

 image by from 15 to 30 minutes' immersion. Striking 



images of the cruciform figures in the nodes of Ranvie: 

 in medullated nerves are obtained by this reagent (S 

 Mayer). Methyl-green. This is chiefly used as a nu 

 clear stain for fresh or recently fixed tissues ; it is alB 1 

 reagent for amyloid degeneration (Heschl), giving 1 

 violet color. Use 0.5 gm. of methyl-green in 20 c.c. of 

 90 per cent, alcohol, 80 c.c. of distilled water, and i 

 c.c. of acetic acid. Stain the tissue for 5 minutes, wa-i 

 in acidulated water, differentiate in 90 per cent, alcohol 

 and dehydrate. The nuclein reaction depends on tin 

 presence of the acetic acid. Arnold recommends a di 

 lute solution of methyl-green containing 0.6 per cent 

 sodium chlorid for staining cells and nuclei. Bizzoz 

 ero has observed that the elements of blood and pus 

 also ciliated epithelium and spermatozoa, do not stair 

 with methyl-green if the cells are highly alkaline ; if 

 the alkalinity is diminished they are dyed violet ; if tin 

 cells are acid, they are colored green. Carnoy regasdi 

 methyl-green as the best stain for nucleoli. Methyl 

 violet. A good chromatin stain. Dissolve 0.5 gja 

 in 200 c.c. of distilled water and 5 c.c. of glacial aceti 

 acid. Stain sections for 20 minutes, wash in distille 

 water, and then in equal parts of glycerin and wau-i 

 Mount in Farrant's medium. This is also a reagO 

 for tissues undergoing amyloid degeneration. Th 

 amyloid substance stains pink. Naphthylamin 

 brown. See Staining of Nerve-tissue, Kaiser 1 

 Method. Nigrosin (Anilin Blue-black). A plad 

 matic stain having a special affinity for ganglion-cell j 

 and much used in the study of the central nervous sv 

 tern. Dissolve 2 gm. in 100 c.c. of distilled wate 

 and stain sections for from 10 to 15 minutes. Orang'i 

 A plasmatic dye used for counter-staining. Dissolve 

 gm. in 20 c.c. of 90 per cent, alcohol and 80 c.c < 

 water. Stain for 10 minutes and wash out in alcohc 

 Phenylene-brown. See Bismarck Brcnun. Quinc. 

 lei'n. See Cyanin. Rubin, Rosei'n. See . 

 Rosanilin Acetate, Sulphate, and Hydrochlorat 

 See Fuchsin. Rose-bengal. Take of rose 

 I gm. , 90 per cent, alcohol 20 c.c, and distilled wal 

 80 c.c. This solution is especially useful for demo 

 strating the beginning of amyloid degeneration ; it stai i 

 the tissues bright-red. It is also recommended 1 

 staining the spinal cord and as a contrast-stain. Si 

 franin. Ffitzner's formula : safranin (Griibler's) 

 part, absolute alcohol 100 parts, water 200 par 

 Flemming uses a concentrated alcoholic solution dilut 

 y^ with water. Babes' formula : (a) equal pa 

 of a concentrated alcoholic and a concentrated 

 solution ; (^) water 100 parts, anilin-oil 2 parts, safrar 

 in excess. The latter may be used according 

 method of Gram, and is recommended for the denu 

 stration of mitotic figures. Solferino. See 

 Spiller's Purple. Rub up in a mortar 2 gm. of S| 

 ler's purple, No 1, with 10 c.c. of alcohol, and 

 IOO c.c. of distilled water. It is used as a doubl 

 and for the fibrin in coagulated blood. If the si 

 to be mounted in balsam, use cedar-oil to clarity 

 Vesuvin. See Bismarck Brown. Victoria Bl" 

 A beautiful nuclear stain. Use a saturated aqueous 

 lution. or 0.25 gm., in 20 c.c. of 90 per cent 

 and 80 c.c. of distilled water. Chromatin and nucl« 

 are stained blue, cytoplasm greenish-blue, or not 

 all. As clove-oil washes out the color, clear 

 oil. This stain has a special affinity for elasti 

 fixation in Flemming's fluid or other clm>in 

 mixture is necessary to secure this reaction. II ( 

 MIN AND COCHINKAI.. These dyes tdve nuclear 

 actions, and are chiefly used for staining in the 111 

 Acetic-acid Carmin. To boiling 45 p 

 acetic acid add carmin until no more will dissolve, ! 

 filter. For use, dilute to I per cent. The con. 



