STAINS, ETC. 



1377 



EXAMINATION MEDIA 



best stained with Loeffler's methylene-blue. If the 

 specimen has been stained with eosin, and the excess 

 washed out, a second or two suffices for the methylene- 

 blue stain. Both oxyphilous and basophilous granules 

 may be stained in the same specimen by preceding 

 the blue stain with eosin. Ehrlick's " Mastzellen .' ' 

 1. In blood these cells are stained by a mixture com- 

 posed of water, loo c.c, absolute alcohol, saturated 

 with dahlia, 50. c.c, glacial acetic acid IO to 1 2. 5 c.c. 

 The leukocytes are stained blue, the granules have a 

 *' metachromic red-violet tint," and correspond to the 

 y-granulations. 2. In tissues : a small piece of fresh 

 tissue, or a section previously hardened in alcohol, is 

 placed in a watch-glass containing anilin-water and 

 20 to 30 drops of a concentrated alcoholic solution of 

 dahlia or gentian. Heat until vapor begins to arise, 

 stain 24 hours, wash in acid-alcohol until nearly de- 

 colorized, dehydrate in absolute alcohol, clear, and 

 mount. Nuclei of the cells are red, the granules in 

 the protoplasm of the granular cells blue. The tissue 

 may also be stained with lithium-carmin. Fixing of 

 Blood. Garlinski ' s Modification of Gaule's Sublimate 

 Method. A small pipet is filled with the indifferent 

 fluid used in counting blood-corpuscles. A drop of 

 blood is drawn into the pipet, where it mixes with 

 the indifferent fluid. A little of this mixture is placed 

 on a slide, and a concentrated watery solution of corro- 

 sive sublimate poured upon it. After a few minutes 

 the morphologic elements of the blood become attached 

 to the glass without alteration of form. The specimen 

 is then washed with water, treated for some minutes 

 with absolute alcohol, and again washed with water, 

 when it is ready for staining. ( Grionlziige der allg. 

 Pathologie d. Zelle. S. M. Lukjanmu, Leipzig, i8gi.) 

 Garlinski's Method. Stain for 2 minutes in Bohm- 

 er's hematoxylin ; wash in I per cent, aqueous solution 

 of alum and distilled water ; then stain in I per 

 cent, aqueous solution of nigrosin for a few seconds ; 

 wash and stain in I per cent, aqueous solution of rose 

 bengal 5 minutes ; wash and stain in anilin-yellow, 

 I per cent, alcoholic watery solution, 5 minutes. 

 Wash, dehydrate, mount in balsam. Cell-protoplasm 

 is yellow ; nuclei are blue or green ; any parasites in 

 the corpuscles are stained by the rose bengal. 

 Hayem's Solution. Used for fixing blood-corpuscles 

 of both animals and man. Dissolve in 200 c.c. of dis- 

 tilled water, 0.5 gm. of corrosive sublimate, 5 gm. of 

 sodium sulphate, and I gm. of sodium chlorid. Run 

 directly from a blood-vessel I part of blood to 100 of 

 the fluid. The corpuscles will be fixed in about 24 

 hours. Decant the supernatant fluid, and wash the 

 corpuscles in water to remove the salts. Hemin 

 Crystals. Place a particle of dried blood on a slide, 

 add a crystal of common salt and two drops of acetic 

 acid, heat over the flame of a spirit-lamp until it steams, 

 and allow it to cool. The crystals may be preserved 

 by removing the acid and mounting them in glycerin- 

 jelly or balsam. Leukocytes. A cover-glass prepa- 

 ration of blood is floated on a solution of eosin, 

 washed and floated on a solution of hematoxylin, 

 washed, dehydrated, and mounted. Methylene-blue 

 or methyl-violet may be used in place of the 

 hematoxylin. Martinotti and Resigotti's Method. 

 Harden small pieces of tissue in absolute alcohol, 

 and color the sections in a water)- solution of safra- 

 nin-0 ; decolorize in 2 parts of a I per cent, solution 

 ->( chromic acid to 8 or 9 parts of alcohol. Wash 

 n absolute alcohol, clear in oil of bergamot, and 

 in balsam. Only the fibrils of the nuclei are 

 -t lined. Rollett's Method of Preparing Hemo- 

 globin Crystals. Defibrinated blood is placed in a plat - 

 num capsule on a freezing mixture, frozen, and then 

 87 



thawed. The lake-colored blood is then poured into a 

 plate until it forms a stratum not more than i^mm. in 

 thickness and allowed to evaporated slowly in a cool 

 place. Sectioning Blood. i.Biondi 's Method, Fix two 

 drops of blood in 5 c.c. of 2 per cent. o»mic-acid solu- 

 tion from one to 24 hours, and then mix the blood and 

 osmium solution with agar-agar jelly melted at 35 to 

 37 C. When cool, harden in 85 per cent, alcohol. 

 After a few days, or when the mass has acquired suffi- 

 cient consistence, embed in paraffin. The sections are 

 treated according to the usual methods, and may be 

 stained with methyl-green, methylene-blue, fuchsin, or 

 safranin ; also, double-stained with methyl-green and 

 eosin. 2. Fod's Method. Coagulated blood or small 

 pieces of hematopoietic organs are fixed in a solution of 

 2 gm. of corrosive sublimate in 100 gm. of Midler's 

 fluid. The Latter fixes the hemoglobin, the sublimate 

 fixes structures of protoplasm and nuclei. Embed 

 in paraffin, section, and stain I to 3 minutes in a mix- 

 ture of Bohmer's hematoxylin 25 gm. , I per cent, 

 aqueous alcoholic solution of safranin 20 gm.,and 

 distilled water 100 gm. Wash in water, then in a 

 weak alcoholic solution of picric acid, dehydrate, and 

 mount in balsam. Weigert's Method for Fibrin. 

 Make celloidin sections, and stain one minute in Weig- 

 ert's fibrin stain : 5 per cent, solution of gentian- vio- 

 let 4.4 c.c, 96 per cent, alcohol 6 c.c, anilin-oil I 

 c.c. Dry with unsized printing paper, and add a drop 

 of Gram's solution saturated with iodin. Most of the 

 stained parts are decolorized. Remove the iodin with 

 printing paper ; clear in equal parts of anilin-oil and 

 xylol, renewing it until all the water is removed. The 

 water gives the section a white appearance. Dry with 

 filter-paper, wash well with xylol, and mount in xylol - 

 balsam. Zenker's Method. For red blood-corpuscles 

 in tissues. The tissue is taken as fresh as possible, 

 placed in Midler's fluid for 24 hours, in which it turns 

 yellow. Longer immersion is detrimental. Wash 

 about two hours in running water ; harden in 50, 70, 

 and 96 per cent., and, lastly, in absolute alcohol. 

 Embed in paraffin. Celloidin sections do not stain as 

 well. Stain on the slide in the Ehrlich-Biondi triple 

 mixture (see Staining Reagents) for 24 hours ; rinse 

 half a minute in running water, and decolorize in 96 

 per cent, alcohol, until clouds of color no longer appear ; 

 dehydrate in absolute alcohol. The red blood-corpus- 

 cles appear a brilliant golden-yellow ; the nuclei of all 

 cells have a violet or green tinge ; the chromatin net- 

 work and nucleoli are invisible. Weigert's fibrin stain 

 may be used in the same way, but not after prolonged 

 immersion in Midler's fluid. (Virch. Arch., 1894, 

 Bd. 135. Folge xiii, Bd. v.) 



EXAMINATION AND PRESERVATION 

 MEDIA. 



Indifferent liquids, glycerin, and resinous preparations 

 used in examining, preserving, and mounting tissues 

 and organisms. I. Indifferent Liquids. Media 

 having a composition and density similar to that of the 

 plasma which constitutes the natural habitat during 

 life of the object they are intended to preserve, and 

 therefore, supposed to have no action on the tissues. 

 To be " indifferent," these liquids must possess such a 

 density and such a proportion of crystalloids and col- 

 loids as will reduce osmotic processes to a minimum. 

 Alum Sea- water. A saturated solution of alum in sea- 

 water is useful for the study and preservation of the tis- 

 sues of marine organisms. Aqueous Humor. This 

 may be obtained from a freshly excised ox's eyeball. 

 Puncture the cornea with a slender, triangular knife, 

 and collect the aqueous humor as it exudes. If only a 

 small quantity is desired, puncture the excised eye of a 



