STAINS 



494 



STAINS 



pyridin. After 15 minutes wash in water, stain with 

 0.25% aqueous solution of anilin blue-black, and then 

 with picrocarmin ; dehydrate, clear in pyridin, mount 

 in balsam diluted with pyridin. Gothard's Method 

 for ganglion-cells: Stain celloidin sections for 24. 

 hours in polychrome methylene-blue and differentiate 

 in a mixture of creasote 5 parts, cajeput oil 4 parts, 

 xylol 5 parts, absolute alcohol 16 parts. Gram's 

 Method. See Staining of Microorganisms, Table of 

 Stains (Illus. Diet.'). I. Giinther's modification: 

 Transfer from the iodin- potassium iodid solution to 

 alcohol, then to a mixture of alcohol, 1 volume, and 

 nitric acid, 3 volumes, and from this again into alco- 

 hol. 2. Nicolle's modification: Decolor in a mixture 

 of alcohol, 2 volumes, and acetone, I volume. 3. 

 Ribbert's modification: Decolor in alcohol containing 

 10 # of acetic acid. Cf. Claudius'' Ale/ hod. Grep- 

 pin's Method for the treatment of Golgi preparations 

 that they may be mounted under a cover: After silver- 

 ing, cut the sections on a freezing microtome, treat 

 them for 30 or 40 seconds with lo$> hydrobromic acid, 

 wash in water, and mount in the usual way. Grim- 

 bert's Medium for the bacillus of Eberth: Make a 

 solution of 2 parts each of amidin, asparagin, neutral 

 potassium phosphate, potassium sulfate, magnesium 

 sulfate, ammonium bimalate, I part each of maltose 

 and magnesium carbonate, in 100 parts of water; add 

 l S% °f g e ' ann > dissolve in a water-bath, cool to 55 

 C. ; add the white of an egg beaten in a little water. 

 Add 5 c.c. of lime-water to each 10 c.c. of the medium, 

 heat in the autoclave at IlO° C. for 15 minutes and 

 filter. Before using add to each tube 1 c.c. of a fresh 

 10% solution of potassium iodid. Gruber and Dur- 

 ham's Method for the agglutination of typhus and 

 cholera bacilli : Place a drop of immunization serum 

 on a cover-glass and beside it a drop of equal size of 

 the culture, as finely divided as possible. Mix and 

 examine on a slide with a ground cell. In doubtful 

 cases put the preparation in the oven for from 15. to 30 

 minutes. Gudden-Weigert Method for medullated 

 nerves: Fix in lofc formalin and harden in alcohol. 

 Treat the sections for 10 hours at room-temperature 

 with 0.5% chromic acid or with \ c / chromic acid 

 heated until it steams; wash and stain in heated 

 "Weigert's hematoxylin acidulated with nitric or hydro- 

 chloric acid. Gulland's Method. 1. lor attaching 

 sections to the slide: Pour a layer of water on a slide 

 and place the sections on the water; heat to 45 or 

 50 C. and the sections will flatten. Remove the 

 excess of water and dry for 24 hours in a thermostat at 

 35 C, finally heating for a moment above the melting- 

 point of the paraffin. Cf. the method of Gaule. 2. 

 For staining blood-cells: Place the fresh cover-glass 

 preparation for from 3 to 4 minutes in a solution com- 

 posed of 25 c.c. of a saturated solution of eosin in 

 absolute alcohol, 25 c.c. of ether, and 5 drops of 

 mercuric chlorid in absolute alcohol (2gm. in 10 c.c); 

 wash in water; stain I minute in saturated aqueous 

 methylene-blue solution; wash in water; absolute 

 alcohol, xylol, balsam. Gum-glycerin: Heat 

 glycerin to boiling and stir in as much powdered gum 

 arable as will dissolve. Wash the object, to remove 

 blood or alcohol, as the case may be, and put it in the 

 gum-glycerin. Keep the object submerged and after 

 24 hours transfer it to 85 % alcohol and shake Vigor- 

 ously at frequent intervals. In a few hours the object 

 will be ready for sectioning. Before staining, wash 

 the sections in water to dissolve out the gum precipi- 

 tated by the alcohol. This is a rapid method suitable 

 for investigations in which histologic differentiation is 

 not important. Gunther's Method of staining bac- 

 teria in blood: Immerse specimen 10 seconds in 5'/ 



aqueous solution of acetic acid until tint of hemoglobin 

 has faded away ; blow off excess of acid and hold speci- 

 men over strong ammonia water to neutralize. Stain in 

 Ehrlich-YVeigert fluid for 24 hours. Decolorize in 

 I : 14 aqueous solution of nitric acid till color fades to 

 light green. Rinse in alcohol, dry. Mount in balsam. 

 Hache's Hematoxylin : Dissolve separately by heat 

 20 gin. of ammonia alum in 200 c.c. of distilled water, 

 and 4 gm. of hematoxylin in 500 c.c. of distilled 

 water ; mix and add a warm saturated solution of 

 sodium bicarbonate ; filter and wash the precipitate for 

 several days, then let it dry at room-temperature. The 

 blue powder thus obtained is soluble in dilute mineral 

 acids, in organic acids, and in a solution of alum. A 

 saturated solution in distilled water containing \ c / f: of 

 glacial acetic acid is recommended for staining nuclei. 

 Let the solution stand for 36 or 48 hours before using. 

 Stain sections for from 12 to 24 hours and differentiate 

 in distilled water. Haffkine's Bouillon for the 

 culture of the bacilli of bubonic plague : Chop a kilo 

 of goat's flesh and heat it at a pressure of 3 atmospheres 

 for 6 hours in dilute hydrochloric acid. Filter, neu- 

 tralize, dilute with water to 3 liters, and sterilize. 

 Haffkine's Prophylactic: Inoculate a flask contain- 

 ing 3 liters of Haffkine's bouillon with a pure culture 

 of pest bacilli ; when the stalactite growth develops 

 shake the flask until the colony sinks to the bottom, 

 and when the growth reappears shake again ; when 

 the stalactite culture forms the third time, heat to 6o° 

 C. for 3 hours. Decant the clear fluid and preserve in 

 hermetically sealed tubes. Dose, I or 2 c.c. injected 

 beneath the skin. Hall's Method for the demonstra- 

 tion of iron in tissue cells : Fix the material for 24 

 hours in a mixture of absolute alcohol 70 c.c, water 

 25 c.c, solution of ammonium sulfate 5 c.c, and 

 harden in graded alcohols from 70^ to absolute. See 

 further ZalewskP 's Method, No. I. Hammar's 

 Method for the study of cleavage in the ova of echino- 

 derms: Fix the ova in a saturated solution of mercuric 

 chlorid in sea-water and stain the sections in Heiden- 

 hain's iron hematoxylin. Hankin's Method for dif- 

 ferentiating pest bacilli : Add 2.5^ of salt to an agar 

 culture. Within 24 hours the bacilli exhibit the in- 

 volution forms that occur in old cultures growing under 

 unfavorable conditions. Hansen's Fuchsin. See 

 Hansen' 1 s Method. Hansen's Hematoxylin : (a) 

 Crystalline hematoxylin, 1 gm., absolute alcohol, 10 

 c.c. (0) Potassium alum, 20 gm., distilled water, 200 

 c.c. Dissolve by heat and filter when cold. (V) 

 Potassium permanganate, I gm., distilled water, 16 

 c.c. After 24 hours mix a ami />, add 3 c.c. of r, and 

 with constant stirring boil one minute. Cool quickly 

 and filter. Hansen's Method for elastin: 

 c.c. of 2% solution of acid fuchsin to 100 c.c. of 

 saturated solution of picric acid; to 9 c.c of this 1 

 ture add one drop of 2% acetic acid. Stain for several 

 minutes or hours; wash in water, each 3 c.c. of which 

 contains 2 drops of the acidified stain. Connective tis- 

 sue, red; elastin and other elements, yellow. Hanz's 

 Stain for gonorrheal secretions : Mix one part ot a 

 saturated solution of fuchsin with 4 parts of a saturated 

 solution of thionin in ." , solution of carbolic 1 

 Hardesty's Method for counting nerve-fibers: Fix 

 the spinal cord and nerves of the frog in situ with 

 osmic acid; wash and treat for an hour with 5', solu- 

 tion of pyrogallic acid. Make transverse sections and 

 photograph them. Harris' Carbol-toluidin : Dis- 

 solve 1 or 2 gm. of toluidm blue in a saturated solution 

 of carbolic acid. Before staining treat the see lions 

 with water; stain for from 5 minutes to 24 hours, wash 

 and differentiate in »iycerin-ether (Grilbler) diluted 15 

 times with water, or in acidulated alcohol ; after from 5 



