STAINS 



491 



STAINS 



Method. (A modification of Krohnthal's.) Harden 

 the tissue in io^r formalin before treating with the 

 formalin- formate mixture. Cut sections without pre- 

 viously embedding and clear in olive oil. Cover- 

 glass Test, split the cover-glasses in two and proceed 

 as in the silk-thread test. Cox's Method I. For 

 neurokeratin : Fix the nerves in I or 2 ^ osmic acid ; 

 wash; dehydrate; clear in bergamot oil, up to 4S 

 hours ; mount in balsam. The bergamot oil dissolves 

 out the myelin and leaves the neurokeratin. 2. For 

 ganglion-cells: Fix for 2 or 3 days in the following 

 mixture : saturate i sublimate solution and 5 % plati- 

 num-chlorid solution 15 volumes each, I f c osmic acid 

 10 volumes, acetic acid 5 volumes; harden in alcohol 

 and embed in paraffin. Place the sections for 8 hours 

 in 25 'r tannin solution, wash, place then for from 5 to 

 10 minutes in 25^ ferric ammonium sulfate solution, 

 after which wash for 10 minutes ; then stain for from 

 12 to 18 hours in Cox's methylene-blue. Cox's 

 Methylene-blue, dissolve I part each of methylene- 

 blue and potassium carbonate in ico parts of water, 

 and shortly before using add 2 ^ phenol solution in the 

 proportion 1:15. Cox-Golgi Method: Treat the 

 tissue for six weeks with Cox's sublimate solution, 

 changing after 24 hours and subsequently once a week. 

 Transfer to 95 fy alcohol for one hour ; to equal parts 

 of alcohol and ether for a half-hour ; to thin celloidin 

 for one hour ; mount in thick celloidin and harden in 

 80^ alcohol for one or at the most two hours. Place 

 the sections in carbol-xylol and mount in balsam under 

 a cover-glas*. Craig's Method for obtaining the 

 flagellated malarial plasmodium : Cleanse the ear or 

 finger, also the slide and cover-glass with alcohol. 

 Make a puncture with a sterile needle and .wipe away 

 the first drops of blood. Gently breathe upon the slide 

 and take up on it the blood from the summit of the 

 second drop and immediately apply the cover-glass. 

 The brief exposure to air and the moisture on the slide 

 are said to hasten flagellation. Cresyl-violet. See 

 Kresyl-vioUt R R. Czaplewski's Stain for bacteria 

 that have been decolored after Gram : Rub up I gm. 

 of fuchsin with 5 c.c. of carbolic acid, and while tritu- 

 rating add 50 c.c. of glycerol and 100 c.c. of water. 

 Darkschewitsch's Method for celloidin serial sec- 

 tions : Fill a beaker of suitable diameter with alcohol ; 

 cut discs of filter-paper of the same diameter, number 

 them, arrange serially, and saturate with alcohol. 

 Gently press a dish against the microtome knife, then 

 strip it off; the sections will adhere to the paper. 

 Preserve the disks, sections uppermost, one above the 

 other in the beaker with alcohol. Deetjen's Method 

 for the investigation of blood platelets : For the study 

 of the vital phenomena mount the platelets in a 

 solution of agar containing sodium chlorid, sodium 

 phosphate, and potassium acid phosphate. For the 

 study of their structure stain with hematoxylin. 

 Diamond's Method for staining Amaba coli : 

 Fix the material in Heidenhain's sublimate salt 

 solution and stain the sections for from 10 minutes 

 to several hours in a mixture of equal parts of 

 carbol-fuchsin and saturated aqueous solution of tol- 

 uidin blue; wash in alcohol. Differentiation, the 

 process of extracting the dye from overstained tissues 

 in the method of indirect or regressive staining. Ex- 

 traction with pure alcohol is termed neutral differentia- 

 tion ; extraction with acidulated alcohol is called acid 

 differentiation. Dimmer's Method for serial cel- 

 loidin sections : Dissolve 16 gm. of gelatin in 300 

 c.c. of warm water ; paint a thin coat of this solution 

 on warmed glass plates. Transfer the sections by 

 tissue paper to the plates and wash them with 70 r 'r 

 alcohol. Absorb the alcohol with bibulous paper and 



press the sections down on the plates. Pour over a 

 photoxylin solution (6 gm. to 100 c.c. of equal parts 

 of absolute alcohol and ether) and when partially dry 

 place the plates in water of from 50 to 55 C. Cut 

 the photoxylin from the edge of the plate, and when 

 the water has dissolved the gelatin the sections, held 

 together by the photoxylin, can be readily separated 

 from the plates and transferred to stains by tissue 

 paper. Dodge's Method for staining starch : Fix 

 in alcohol and (if necessary) bleach the chlorophyl 

 bodies with javelle water. Stain with iodin potassium 

 iodid solution (1 : 2:300), wash with distilled water, 

 and treat with I <£ solution of silver nitrate in a bright 

 light for a few minutes. Reduce in hydrochinone 2 

 gm., sodium sulfite IO gm. , water 100 c.c. , and I drop 

 of 10 J?- solution of potassium carbonate to each I c.c. 

 Dollken's Method for staining very young brains. 

 Cut sections from 30// to 50 ft thick, attach them to 

 the slide after Obregia (see Fixath-es, Table of Stains 

 [Illus. Diet] ), and stain for4 or 5 days in cold hema- 

 toxylin, then at 37 C. for 2 hours; when cool treat 

 with spring- water for 5 hours, then for 1 5 minutes 

 with distilled water containing 2 or 3 drops of potash so- 

 lution to the liter; differentiate in o. 5O solution of 

 potassium permanganate ; when the gray tissue becomes 

 transparent wash in distilled water and treat with 1 

 lution of oxalic acid until the gray tissue turns light 

 brown. The material should be fixed in chrcmic acid or 

 in J'f formaldehyd. Doutrelepont's Method for 

 the bacilli of syphilis : Stain for 24 hours in I <£ solution 

 of methyl-violet, decolor for a few seconds in dilute 

 nitric acid, and transfer to 60 & alcohol for 10 minutes; 

 stain for a few minutes in aqueous solution of safranin 

 and wash in 60% alcohol. Driesch's Method for 

 the study of pressure phenomena in the dividing 

 eggs of sea-urchins. Three minutes after fertiliza- 

 tion shake the eggs in a test-tube with water, for 

 a few seconds, to rupture and detach the fertiliza- 

 tion membrane. Transfer the eggs to a slide. Ehr- 

 lich's Iodin Method : Stain the fixed film in a 

 syrupy solution of gum arabic containing I f f of 

 Lugol's solution : Leukocytes stained brown indicate 

 a suppurative process. Ehrlich's Stains for the 

 granules of leukocytes : 1. Acidephilous or ecsincphil- 

 cus mixture. Two parts each of indulin, aurantia, 

 and eosin ; glycerol, 30 parts. Suitable for staining 

 sections and cover-glass preparations. This is also 

 known as " Mixture C." 2. "Triadd" mixture. 

 Dissolve (a) I gm. of orange-yellow (extra) in 50 c.c. 

 of distilled water; (b) I gm. of acid fuchsin extra in 

 50 c.c. of distilled water ; (r) I gm. of crystalline 

 methyl-green in 50 c.c. of distilled water. Let the 

 solutions settle. Then mix 1 1 c.c. of solution a with 

 IO c.c. of solution b ; add 20 c.c. of distilled water 

 and 10 c.c. of absolute alcohol ; to this mixture add a 

 mixture of 13 c.c. of solution c, 10 c.c. of distilled 

 water, and 3 c.c. of absolute alcohol. " Let the stair, 

 stand for one or two weeks before unng. Ehrlicb- 

 Lazarus Method for the basophil granules of mast- 

 cells : Use kresyl-violet. See the method of Biel- 

 schowsky and Plien. Ehrlich- Weigert Method for 

 staining tubercle bacilli: Prepare a mixture of 1.1 

 parts of a saturated alcoholic solution of methyl violet, 

 I part of absolute alcohol, and 10 parts of anilin water. 

 In this stain the film for from 2 to 5 minutes, heating 

 until it steams ; decolor for a few seconds in nitric 

 acid diluted with 3 volumes of water. Mash in 6o<{f 

 alcohol, then in water. Counterstain for 5 minutes 

 in a saturated aqueous solution of vesuvin. Eisen's 

 Method. I. For attaching sections to the slide. 

 Flood the slide with 80^ alcohol, place the sections 

 on the liquid, and put them in the oven, at 55 C. 



