STAINS 



493 



STAINS 



anilin for a few minutes, then with oil of lavender 

 and xylol, and mount in balsam. Fluorescein, an 

 acid dye used in alcoholic solution as a differentiating 

 and bleaching medium and as a stain in the following 

 procedure : Dissolve 5 g m - each of fluorescein and 

 sodium carbonate in 30 c.c. distilled water. Inject 1 

 c.c. under the skin of a narcotized animal ; when the 

 skin plainly shows the color, kill the animal. The 

 dve appears first in the bloodvessels, later in the lymph- 

 atics. Foa's Mixture : Dissolve 2 gm. of subli- 

 mate in iooc.c. of Miiller's fluid heated nearly to boil- 

 ing. Fix hematopoietic tissue from 2 to 24 hours. 

 >h in alcohol. Frankel's Method for the dem- 

 onstration of moulds : Tease the material in 50^ 

 alcohol containing a few drops of ammonia and mount 

 in glycerol ; or stain with vesuvin or methylene-blue 

 and mount in balsam. Frankel-Gabbet Method 

 for tubercle bacilli : The dried and fixed preparation 

 is placed for 10 minutes in a solution consisting of 

 fuchsin 1 part, alcohol 10 parts, carbolic acid 5 parts, 

 distilled water loo parts, then dried with filter-paper 

 and placed for 5 minutes in a second solution of methyl- 

 ene-blue 2 parts, sulfuric acid 25 parts, distilled water 

 100 parts ; it is then washed and dried. If the stain 

 has been successful, the preparation will have a faint 

 blue color. Freeborn's Mixture for clearing tissues: 

 Mix I part of oil of origanum cretici and 3 parts of oil 

 of thyme and add a large quantity of powdered chalk ; 

 after 24 hours filter. Fuchs' Method for the study 

 of sputum: Fix by heat, stain for 2 minutes in 0.5% 

 alcoholic solution of eosin, and decolor in 50^ alco- 

 hol. Counterstain with methylene-blue. Futcher- 

 Lazear Method for the malarial parasite : Fix the 

 film for one minute in a mixture of 10 c.c. of 95 > 

 alcohol and 2 drops of formalin ; wash, dry, and stain 

 for 15 seconds in carbol-thionin, prepared by mixing 

 20 c.c. of a saturated solution of the dye in S°% alco- 

 hol and loo c.c. of 2% aqueous solution of carbolic 

 acid. Gad's Method for nerve-endings in striped 

 muscle and bloodvessels : Place small muscle-bundles 

 for 18 hours in a mixture of acetic acid I part, glycerol 

 I part, I % aqueous solution of chloral 6 parts ; tease 

 in glycerol and stain for from 3 to 10 days in Ehrlich's 

 hematoxylin I part, glycerol I part, 1% aqueous solu- 

 tion of chloral 6 parts ; mount in acidulated glycerol. 

 Galli's Method for neurokeratin: Fix a sciatic nerve 

 for 20 minutes in Miiller's fluid; tease it and place it 

 for 2 days in Miiller's fluid diluted with 2 parts of 

 water; transfer into glycerol (containing a drop of 

 glacial acetic acid to each cubic centimeter) for 15 

 minutes and, without washing, stain for 20 minutes in 

 aqueous solution of china blue. Wash in alcohol ; 

 turpentine; balsam. Garcia Rigo's Method of 

 rapid double staining for blood examination : A drop 

 of blood on a cover-glass is diluted with a drop of 

 simple bouillon (kept sterile with a little formol) and 

 the two stirred with a sterile platinum wire until mixed. 

 The cover-glass resting on the end of a slide is then 

 warmed over an alcohol flame for less than a minute. 

 Eosin stain is next used and washed with water ; then 

 methylene-blue and washed again. The specimen is 

 then dried and mounted in Canada balsam, the whole 

 process occupying 5 minutes. Gatehouse's Method 

 for staining embryonic tissues and for restoring faded 

 slides : Saturate filtered turpentine with picric acid and 

 cautiously add crystals of iodin until the yellow color 

 has a brown tint. Gautier's for blood. Followed 

 Romanowsky' s technic. Employed for the methylene- 

 blue solution: Methylen-blau (Badische Soda-anilin 

 fabrik), Marke C orBGX; For the eosin solution: 

 eosin (Badische Soda-anilin fabrik), Marke A. 

 Gelpke-Weigert Method for pathologic nerves: 



For transverse sections of atrophied nerves dilute the 

 differentiating fluid with 50 volumes of water and im- 

 merse for 12 hours. For longitudinal sections, dilute 

 with 10 volumes of water. The process is applicable 

 to tissue hardened in alcohol or other fluid, provided 

 it is treated with a solution of a chromic salt until it 

 becomes brown, before mordanting in the copper or 

 cyanid solution. See further ll'tigert's Alethoa, Table 

 of Stains (Illus. Diet. ). Gilson's Method. A rapid 

 celloidin process. Dehydrate the object, soak it in 

 ether, and put it into a test-tube with thin celloidin 

 solution. Place the tube in melted paraffin and boil 

 until the cellodin is of a syrupy consistence. Mount 

 on a block of hardened celloidin and harden in chloro- 

 form or in a mixture of chloroform and cedar-oil. In 

 cutting use cedar-oil to wet the knife and the object. 

 Gilson's Mixture : Nitric acid (sp. gr. 1. 456) 78 c.c, 

 glacial acetic acid 22 c.c, mercuric chlorid 95 to 100 

 gm., 6of c alcohol 500 c.c, distilled water 4400 c.c. 

 A generally useful fixing medium. \\ hen used for 

 marine animals, add a few crystals of iodin. Gly- 

 cerin-ether. See L'tnta's Method (7). Glychema- 

 lum, hematein 0.4 gm. (rubbed with a few drops of 

 glycerol until it dissolves), alum 5 gm., glycerol 30 

 c.c, distilled water 70 c.c. (Mayer). Godlewski's 

 Method for the study of developing striped muscle- 

 fibers : Fix salamander larva- and the extremities of 

 infant mice or guineapigs in saturated aqueous solution 

 of sublimate containing 2% of acetic acid and harden 

 in alcohol. Embed in paraffin and cut longitudinal, 

 transverse, and oblique 5 u sections ; stain in thionin or 

 hematoxylin and afterstain in eosin. S., Goldhom's, 

 for blood. Preparation of the solution of polychrome 

 methylene-blue. Solution A. — (Merck's medicinal 

 methylene-b'.ue : Grubler's methylene blue rectified, 

 and methylene-blue [Koch]). Dissolve 2 gm. meth- 

 ylene-blue in 300 c.c. warm water. Add to this 4 gm. 

 lithium carbonate, shaking constantly. Heat in an 

 evaporating dish on a water-bath, the water touching 

 the dish. Stir the solution occasionally. Remove in 

 1 5 to 20 minutes. Do not filter. Set aside for several 

 days. Then add dilute acetic acid ($?c) until the 

 solution is only faintly alkaline. Solution B. — A 

 o. 1 % aqueous solution of eosin. Fix blood-films in 

 methyl alcohol for 15 seconds. Wash in running water. 

 Stain in Solution B for 7 to 30 seconds. Wash. Stain 

 in Solution A for 30 seconds to 2 minutes. Wash 

 thoroughly in running water. Dry by agitating in air, 

 not between filter-paper. The eosin may be added to 

 the methyl alcohol (enough to make ao.iJJ solution); 

 or Solution B may be added to Solution A (1 :4), but 

 this easily produces a precipitate (the neutral stain). 

 These give good results. Mixtures of methyl alcohol, 

 eosin, and polychrome methylene-blue give poor 

 results. The depth of the chromatin stain depends on 

 the length of staining. To stain the chromatin of 

 half-grown malarial parasites. i' 2 ' to 2 minutes are 

 necessary, while the chromatin of the hyaline forms 

 stains in 10 seconds. Repeated staining may improve 

 the chromatin violet. To do this the blood-film may 

 be stained with Solution B 5 seconds, with Solution A 

 10 seconds. Golgi's Method for the restoration of 

 overhardened tissue : Wash in a half-saturated solution 

 of copper acetate until it yields no precipitate and 

 return for 5 or 6 days to the osmium-bichromate 

 mixture. The tissue will then take the silver and the 

 sections can be mounted in thickened cedar oil under a 

 cover-glass. Golgi's Mixture : Potassium bichromate 

 (3.5^ solution) 54 c.c, osmic aci : . c.c. 



Goodall's Method for the spinal cord : Cut the fresh 

 tissue on a freezing microtome ; float the sections on 

 water; as soon as possible, drain and float them on 



