STAINS 



509 



STAINS 



potash I gm., alcohol loo c.c.) ; when the sections 

 become a transparent brown-red wash for 5 minutes in 

 distilled water ; counterstain for from 15 to 30 minutes 

 in saturated solution of safranin diluted with an equal 

 volume of water ; wash and dehydrate in absolute 

 alcohol. Stroschein's Method for the sedimentation 

 of sputum: Agitate thoroughly in a test-tube 5 c.c. 

 of sputum with from 5 to 15 c.c. (according to the 

 consistence of the secretion) of a mixture of borax- 

 boric acid solution and water in the proportion of 1 : 3. 

 Sedimentation occurs in from 24 to 48 hours. Sub- 

 stantive Staining, a histologic stain obtained by 

 direct absorption of the pigment from the solution in 

 which the tissue is immersed. Dyes that combine 

 directly with the substance acted on are called sub- 

 stantives dyes. Cf. Adjective Staining. Subtractive 

 Staining, a so-called theory of Heidenhain's, based on 

 the hypothesis that a general stain satisfies the affinities 

 of some cell structures, that hold it in subsequent treat- 

 ment with specific dyes, while the other structures give 

 up the general stain and then take the specific stain. 

 Sudan III, a selective stain for fat. Prepare a satu- 

 rated solution in 95 % alcohol, dilute two-thirds with 

 50% alcohol, and filter. Stain sections for from 5 to 

 10 minutes, wash for about the same time in 60 or 70$& 

 alcohol and mount in glycerol. Small oil drops yel- 

 low, large ones orange. For staining the fat granules 

 in the elements of tissues undergoing fatty degenera- 

 tion use the undiluted stain. The tissue may be fixed 

 in Midler's fluid or cut fresh on the freezing microtome. 

 Symington's Method for showing the relation of the 

 skull to the external and internal parts of the brain : 

 Harden the entire head by repeated injections of for- 

 malin, through the carotid and vertebral arteries. In- 

 ject a solution of gum, fix in a box filled with the gum 

 solution, freeze, and make sections. Tandler's 

 Method for celloidin sections : Transfer the sections 

 from the knife to the slide, mop up the surplus alco- 

 hol, and cover the sections with a strip of paper having 

 twice the length of the slide ; turn the face end round 

 against the under surface and against this place an- 

 other slide. Put the prepared slides, one above the 

 other, in a dish with water or alcohol. Tanzer's 

 Orcein. Orcein 0.5 gm., absolute alcohol 40 c.c, 

 distilled water 20 c.c, hydrochloric acid 10 drops. 

 Tellyesnicky's Fluid. Potassium bichromate 3 

 gm., glacial acetic acid 5 c.c, water 100 c.c The 

 time for fixing is from one to two days, according to 

 the size of the object. Wash in water and harden in 

 alcohol. Thalmann's Medium for the cultivation 

 of gonococci : Sterilize horse's brain in a steam ster- 

 ilizer for one hour; divide it into thin slices, put them 

 into petri dishes, and sterilize twice, a half-hour each 

 time. Thionin, the uses and technic are the same as 

 for methylene-blue. A saturated solution in 50 % 

 alcohol diluted with 5 volumes of water is used for 

 staining. Cf. the methods of Lenhossek and Harris. 

 Thorn's Method for staining goblet cells: Harden 

 the tissue in alcohol and stain the sections for 15 min- 

 utes in Mayer's hematin ; wash in 70% alcohol and 

 stain for a very short time in a solution of bismarck 

 brown in 70J& alcohol. The cells containing mucus 

 are brown. Thoma's Method for the numeration 

 of leukocytes : Dilute the blood in the proportion of 

 I : IO with water containing 0.3^ anhydrous acetic 

 acid. This dissolves the colored blood-cells. Tim- 

 berlake's Fluid for fixing kinetic nuclei in plant 

 cells: 1. Iridium chlorid 0.5 gm., water 100 c.c, 

 glacial acetic acid I c.c. 2. A I c ' ( solution of iridium 

 chlorid with 3^ of acetic acid. Tinctorial Preoc- 

 cupation, a theory of staining formulated by Unna 

 and identical with subtractive staining. Toluidin 



Blue, a regressive anilin dye resembling methyl- 

 ene-blue. See Harris Carbol-toluidin. Touton's 

 Method for gonococci in tissue sections : Stain in 

 carbol-fuchsin and wash in alcohol. Trambusti's 

 Method for blood in tissue sections : Fix the object 

 in Flemming's mixture and place the sections for 24 

 hours in 1% solution of thionin in anilin water 

 (4 : 100) ; treat with acid alcohol and stain in aqueous 

 solution of eosin, then in alcoholic solution of eosin. 

 Mount in xylol balsam. Turner's Method for the 

 study of nerve-cells: Place a thin slice of gray 

 nerve tissue in 0.5^ solution of methylene-blue. 

 After 12 hours transfer a very minute fragment to a 

 slide; add a drop of Farrant's medium, and apply a 

 cover-glass. Crush the tissue by careful pressure on the 

 cover ; this should be done under the microscope. 

 Cf. Vincenzi 1 s Method. Unger's Method for the 

 study of mammary glands : Fix very small pieces 

 from 2 to 5 days in Midler's fluid and then in a mix- 

 ture of 2 parts of Miiller's fluid and I part of \f c os- 

 mic acid, renewed daily ; wash in water, harden for 3 

 days in absolute alcohol, and embed. The entire pro- 

 cess should be done in the dark. The sections may be 

 treated for a day with 20^ formic acid and counter- 

 stained with safranin. Fixation in boiling water or 

 alcohol also gives good results. Mount in colopho- 

 nium benzine. Unger's Methyl-green. Methyl- 

 green, from 0.15 gm. to 0.3 gm., water ico c.c, 

 hydrochloric acid 3 drops. This liquid is recom- 

 mended in particular for the staining of spermatozoa. 

 The dry cover-glass preparation should be treated with 

 the stain for several hours. Unna's Hematoxylin, 

 a constant half-ripe stock solution. Hematoxylin I 

 gm., alum logm., alcohol 1 00 c.c , water 200 c.c, sub- 

 limed sulfur 2 cc. If the sulfur be added 2 or 3 

 days after preparing the hematoxylin solution, it will 

 arrest oxidation and the stain will be ready for use at 

 this stage. The oxidation of alum-hematoxylin solu- 

 tions can be instantaneously accomplished by adding a 

 litde neutralized hydrogen dioxid. See Harris' Hem- 

 atoxylin. Unna's Method for collagen : 1. Stain 

 sections of alcohol material for 5 minutes in strong so- 

 lution of polychrome methylene blue, then for 15 

 minutes in neutral I % solution of orcein in absolute 

 alcohol ; wash in alcohol ; bergamot ; balsam. Col- 

 lagen dark red ; nuclei blue ; granules of mast-cells 

 carmin red ; cytoplasm of plasma-cells blue. 2. 

 Stain sections for 20 seconds in I r ( solution of water- 

 blue ( Wasserblau ) ; wash, and stain for 5 minutes 

 in neutral aqueous I % solution of safranin ; wash in 

 water and then treat with absolute alcohol until the 

 blue color reappears ; collagen sky-blue ; nuclei red ; 

 cytoplasm violet. 3. For collagen, elastin. and smooth 

 muscle. Stain with hot orcein (see Staining Reagents) 

 for IO minutes, wash in dilute alcohol, stain with 

 hematein for 10 minutes, and treat for a few seconds 

 with acid alcohol ; wash, and place in a 2 a r solution 

 of acid fuchsin for 5 minutes, in saturated aqueous 

 solution of picric acid for 2 minutes, then in satu- 

 rated alcoholic solution of picric acid for 2 minutes; 

 absolute alcohol ; oil ; balsam. Elastin brown-red ; 

 collagen red; muscle-fibers yellow with gray-violet 

 nuclei. 4. For elastin and smooth muscle. Stain as 

 in 3, substituting polychrome methylene-blue for hem- 

 atein and I fy potassium permanganate for the acid 

 alcohol. Elastin brown-red ; collagen decolored ; 

 muscle fibers violet. 5. For smooth muscle. Stain 

 sections for 10 minutes in polychrome methylene- 

 blue ; wash, and fix in I # red prussiate of potash ; 

 differentiate in acid alcohol for 10 minutes ; absolute 

 alcohol ; oil ; balsam. The collagen is decolored. 

 6. For keratohyalin. Overstain in hematoxylin, treat 



