STAINS 



495 



STAINS 



to 15 minutes wash in alcohol. Eosin in alcohol may 

 be used as a counterstain. In this case omit the differ- 

 entiation and stain for from a half to 2 minutes and 

 wash in alcohol. Harris' Hematoxylin : Dissolve 

 1 gin. of hematoxylin in 10 c.c. of alcohol and add to 

 200 c.c. of a saturated aqueous solution of alum; heat 

 to boiling and add 0.5 gm. of mercuric oxid ; when 

 the solution turns a dark purple, remove from the flame 

 and cool quickly. For use dilute to the color of port- 

 wine with aqueous solution of alum. Harris' Method. 

 1. For staining pest bacilli in tissue sections: Stain for 

 4$ hours in Harris' carbol-toluidin and differentiate in 

 glycerin-ether. 2. For amyloid substance : Stain the 

 sections of material rixed in alcohol material for 24 

 hours in carbol-toluidin blue ; wash in water; mordant 

 for 2 hours in saturated solution of red or yellow prus- 

 siate of potash or ammonium molybdate. Mount in 

 balsam. Amyloid, red; remaining tissue, various 

 shades of blue. 3. For tissues stained in methylene- 

 blue : Wash in water and place the object in a saturated 

 solution of potassium ferrocyanid ( or ferricyanid | cooled 

 nearly to zero, adding a trace of osmic acid to prevent 

 maceration ; after from 3 to 24 hours wash for one 

 hour in distilled water, dehydrate in ice-cold absolute 

 alcohol, clear- xylol or cedar oil, and embed in paraffin. 

 4. For elastin : Stain sections from 5 to 10 minutes in 

 Harris' hematoxylin and wash for a minute in I % 

 nitric acid in alcohol. Haug's Method for fixing and 

 decalcifying very delicate objects. Prepare a mixture 

 of 1^ osmic acid IO c.c, 1 <£ chromic acid 25 c.c, 

 distilled water 65 c.c. Subsequently wash in water 

 and harden in 70% alcohol. Hauser's Method for 

 sections of gelatin cultures of bacteria : Moisten the 

 cotton- wool plug of the thrust culture or the fiber-paper 

 of the plate-culture with a few drops of formalin and 

 place the whole in a covered vessel with a watch-glass 

 containing a wad of cotton moistened with 10 or 15 

 drops of formalin. The gelatin attains the consistence 

 of celloidin that has been hardened in alcohol and can 

 be cut on the microtome. Hayer's Method for in- 

 f.isoria: The entire process is carried out in a glass 

 cylinder (5 cm. long and 7 mm. wide) open at both 

 ends, with a piece of parchment paper tied over one 

 of the openings. By removing the parchment the 

 paraffin can be pushed out in the for 11 of a cylinder 

 with the embedded objects at that end of it. Heiden- 

 hain's Fluid: Saturate hot 0.5^ sodium chlorid solu- 

 tion with mercuric chlorid. Held's Fluid : Mercuric 

 chlorid I gm., 4% acetone 100 c.c. After fixation 

 wash in acetone gradually increased in strength. 

 Held's Method for ganglion cells: Stain sections for 

 I or 2 minutes in a warm solution of erythrosin, 1 gm., 

 in 150 c.c. of water and 2 drops of glacial acetic acid; 

 wash in water and stain in a mixture of equal parts of 

 NissPs methylene-blue and 5^ acetone, wanning until 

 the odor of acetone disappears. When cool differen- 

 tiate in o. I ^ solution of alum, wash in water, and 

 dehydrate in alcohol. Helianthin. The same as 

 Water Blue. Heller's Method. 1. For the osmica- 

 tion of medullated nerve-fibers : Harden the tissue in 

 Miiller's fluid. Stain the sections in I % osmic acid, 

 in the oven for IO minutes, at room-temperature for a 

 half-hour; wash in water; reduce in 5<£ pyrogallic 

 acid for a half-hour, oxidize in 2.5 V potassium per- 

 manganate for from 3 to 5 minutes, decolor it 

 oxalic acid for from 3 to 5 minutes. 2. For mounting 

 objects for sectioning : Pin a piece of paper about the 

 cork or block so that it projects and forms a trough into 

 which the celloidin can be poured around the object. 

 Harden in the vapor of alcohol by suspension in a 

 closed cylinder containing a few centimeters of alcohol. 

 Hemosiderin, amorphous yellow to black-brown iron- 



containing fragments occurring in thrombi or hemoi- 

 rhagic infarcts. In sections of material hardened in 

 alcohol or formalin, treated for a few minutes with a 

 . meous solution of potassium ferrocyanid and ex- 

 amined in glycerol containing 0.5^ of^ hydrochloric 

 acid the pigment appears in the form of dark blue 

 granules. Henking's Dahlia and Osmic Acid : 

 Dahlia 0.04 gm. , \' ;C osmic acid 1 c.c, formic acid 3 

 c.c", glycerol 16 c.c, distilled water 80 c.c. For its 

 use see Henking'' s Method. Henking's Method for 

 the ova of insects: Tease them in a drop of Henk- 

 ing's dahlia and osmium mixture and examine. To 

 preserve the preparation simply lute the cover-glass. 

 Henneguy's Method. 1. For the study of mitosis : 

 Treat sections for 5 minutes with I 'Jc potassium per- 

 manganate solution ; wash in water and stain in 

 safranin ; wash in alcohol. Karyoplasm and achroma- 

 tin, spindle gray; chromosomes and nuclear membrane, 

 brilliant red ; astrophere and centrosome, less intenseiv 

 stained. 2. For fixing sections to the slide : Spread 

 a film of Mayer's albumen on the slide and over the 

 albumen a drop of water ; on this arrange the sections 

 and warm, but not to the melting-point of the paraffin ; 

 when the sections are flat, evaporate the water at 40° 

 C. Hermann's Method for the study of mitosis: 

 Stain tests of proteus 12 to 18 hours in the dark with 

 the following solution: hematoxylin I gm., water 30 

 c.c, absolute alcohol 70 c.c. ; treat for the same time, 

 in the dark, with 70% alcohol. Embed and treat the 

 sections with pale rose-colored solution of potassium 

 permanganate until they become ochre-color; rinse in 

 water and decolor in Pal's oxalic-acid mixture (see 

 Table of Stains) diluted with 5 volumes of water; 

 stain 3 to 5 minutes with safranin. Herrick's Method 

 for embedding tissue impregnated with methylene- 

 blue : Treat the object with glycerin and then place it 

 for a day in a mixture of glycerin and gum arabic 

 Transfer to a paper tray and leave it exposed to the air 

 until by evaporation it has attained a consistency suit- 

 able for sectioning. Hertwig's Method for determin- 

 ing the influence of temperature on the fertilization of 

 the ova of the chick, frog, and sea-urchin : Cool the 

 eggs for a half-hour or heat them above 31 ° C. and 

 over-fertilization takes place. In further cooling or 

 heating no fertilization occurs. Cf. fertilization, 

 Polyspermous. His' Medium for the differential 

 culture of the typhoid bacillus. I. The tube culture- 

 medium : triturate 5 gm. of agar, 80 gm. of gelatin, 5 

 gm. of beef extract, and 5 gm. of salt ; add a liter of 

 water and enough hydrochloric acid or soda solution to 

 produce a reaction of 1.5^ of normal acid, using 

 phenolphthaleid as the indicator. Clear with I or 2 

 eggs beaten in 25 c.c. of water, add 10 gm. of glucose, 

 boil for 25 minutes, and filter through absorbent cot- 

 ton. 2. For the plate culture use 10 gm. of agar, 25 

 gm. of gelatin, 5 gm. each of beef extract and salt, and 

 IO gm. of glucose. The medium must contain not less 

 than 2 <'r of normal acid. The typhoid bacillus alone 

 has the power of clouding these media. Hodenpyl's 

 Method for attaching sections to the slide or cover- 

 glass: Add to 150 cc of distilled water 50 c.c. of 

 white of egg and 50 c.c. of a solution of salicylic acid 

 that has been slightly alkalized by lithium carbonate. 

 Soak the sections in this for 2 or 3 minutes and then 

 place them on a cover-glass. Hofbauer's Method 

 for staining the iodinophil granules of leukocytes: 

 Dry the film and stain I minute in a solution of iodin, 

 I part, potassium iodid. 3 parts, and water. 100 parts, 

 brought to a syrupy consistence by the addition of gum 

 arabic. Remove the excess of the stain with filter- 

 paper, to prevent diffuse coloring. Cf. Ehrlich's 

 f. Hoffman's Method. 1. For the demon- 



