STAINS, ETC. 



1400 



STAINING REAGENTS 



to be better than alum-carmin for staining in toto. 

 Mayer's Paracarmin. Dissolve carminic acid I gm., 

 aluminum chlorid o. 5 gm. , calcium chlorid 4 gm. in 

 100 c.c. of 7° per cent, alcohol, with or without heat. 

 Filter, after precipitation, and the solution will have a 

 clear-red color. Suitable for staining bulky objects 

 with large cavities, such as Salpa. Orth's Carmin 

 Stains. Lithium-carmin. Digest 2.5 gm. of carmin 

 in 97.5 c.c. of a saturated solution of lithium carbonate 

 and filter. The stain is diffuse, but on treatment with 

 acid alcohol becomes restricted to the nuclei. Tissues 

 refusing to stain in any other medium do so in this 

 fluid. Picrolithium-ca?'imn consists of I part of 

 lithium carmin and 2 or 3 parts of a saturated solution 

 of picric acid. After staining, wash out in acid alcohol. 

 Partsch's Cochineal Alum-carmin. Boil powdered 

 cochineal in 5 per cent, alum solution, filter, and add a 

 little salicylic acid as a preservative. This is said to 

 give more delicate differentiation than other alum- 

 carmins. Partsch-Grenacher's Alum-carmin. A 

 nuclear stain, prepared as follows : Boil for 15 minutes 

 2 gm. of pure carmin, No. 40, and 5 gm. of ammonium- 

 alum in 200 c.c. of distilled water. Cool, filter, and 

 add 2 c.c. of carbolic acid. Ranvier's Carmin. This 

 is composed of carmin I gm., ammonia I c.c, water 

 100 c.c. Rub up the carmin in a mortar with a little 

 water and add the ammonia. When the carmin is all 

 dissolved add the rest of the water. If there is an 

 excess of ammonia, heat the solution until the carmin 

 begins to precipitate. Reeves' [J. E.) Borax-car- 

 min. A nuclear stain, consisting of the best No. 40 

 carmin 4 gm., pulverized borax 8 gm. , and carbolized 

 water, 2 per cent., 120 c.c. Heat in a porcelain vessel 

 to boiling, and, when cool, filter. Stains in from I 

 minute to }/% an hour, according to the character of 

 the tissue. A decolorizing and differentiating fluid 

 composed of pure hydrochloric acid 10 c.c, 70 per 

 cent, alcohol 105 c.c. may be used with this stain. 

 After immersion in this fluid the section must be well 

 washed with water, to remove the acid, and if desired 

 it may now be double-stained in a weak, aqueous solution 

 of sodium sulph-indigotate. Rollet's Acid Carmin. 

 Boil pulverized carmin in dilute sulphuric acid, filter 

 off the red precipitate, and dissolve in water for use. 

 Schneider's Acid Carmin. Boil pulverized carmin 

 in 45 per cent, acetic acid until no more will dissolve, 

 filter, and dilute to I per cent, for use. Schweigger- 

 Seidel's Acid Carmin. Add an excess of acetic acid 

 to ammonium-carmin, and filter. Sodium Carminate. 

 See Staining of Nerve-tissue, Schmaus' 1 Method. 

 III. Hematoxylin and HemateTn. Solutions of these 

 substances are used for staining sections, and also tis- 

 sues, in toto, especially such as have been fixed in chrom- 

 ium and osmium fluids. Bohmer's Hematoxylin. A 

 valuable nuclear stain. Dissolve I gm. of hematoxylin 

 in 100 c.c. of absolute alcohol. Make a second solu- 

 tion of 5 gm. of alum in 100 c.c. of distilled water. 

 Add the first solution, drop by drop, to the second, 

 until a deep- violet color is obtained. Let the mixture 

 stand in an open vessel, exposed to the light, for 

 several days. Filter, and add a little thymol to the 

 filtrate. Cook's {Alleyer) Hematoxylin. An alum- 

 hematoxylin containing copper sulphate. Reduce in 

 a mortar 6 grams each of extract of logwood and 

 alum and I gram of copper sulphate, and dissolve in 

 40 c.c. of water. Filter after 2 days, and add a 

 crystal of thymol to the filtrate. Cuccati's Iodin- 

 hematoxylin. Dissolve 25 gm. of chemically pure 

 potassium iodid in 25 c.c. of distilled water. Pour 

 this solution, slowly and with constant agitation, into 

 75 c.c. of absolute alcohol, and keep in a well-stoppered 

 bottle. Rub up in a mortar 75 eg. of crystallized hem- 



atoxylin with 6 gm. of chemically pure, neutral pota 

 alum, and add 3 c.c. of the iodin-solution ; keep the 

 mixture agitated, and gradually add the remainder of the 

 solution. Shake for some time, to dissolve the alum, stand 

 aside for from 10 to 15 hours, shake again, filter, and 

 preserve in a tightly stoppered bottle. Objects should 

 be left in the liquid for 10 hours, then washed, dehy- 

 drated, and mounted. The solution is said to be a pure 

 chromatin stain. Delafield's Hematoxylin. An 

 excellent nuclear stain. Dissolve 4 gm. of hematoxy- 

 lin in 25 c.c. of absolute alcohol, and add 400 c.c. of 

 a saturated, aqueous solution of ammonium-alum. Ex- 

 pose to light and air for 3 or 4 days ; filter ; add to the 

 filtrate 100 c.c. each of glycerin and methyl-alcohol. 

 Ehrlich's Acid Hematoxylin. Used for staining 

 sections and in the mass. Dissolve I gm. of hematox- 

 ylin in 30 c.c. of alcohol, and add 5° c - c - eacn of 

 glycerin and water, alum in excess, and 4 c.c of gla- j 

 cial acetic acid. Let the mixture ripen in the light un- i 

 til it acquires a deep-red color. Objects stained in it 

 should be washed in undistilled water. Ehrlich's Am- 

 moniated Hematoxylin. Dissolve 2 gm. of hema- 

 toxylin and 0.4 gm. of ammonium carbonate in 80 c.c. 

 of 90 per cent, alcohol. Expose in an evaporating dish 

 for 24 hours, warm, and bring up to 40 c.c. by adding: 

 50 per cent, alcohol, and then add 2 gm. of ammonium-! 

 alum, 80 c.c. of distilled water, 100 c.c. of glycerin, 

 and 10 c.c. of acetic acid. Dilute 1 : 10 for use. 

 Gage's Hematoxylin. Boil 7.5 gm. of alum in 20c 1 

 c.c. of distilled water for about 5 minutes; when cool.! 

 add enough freshly boiled distilled water to make up j 

 the loss by evaporation; add 4 gm. of chloral, and then 

 o.l gm. of hematoxylin dissolved in 10 c.c. of 95 perl 

 cent, alcohol. Let the mixture ripen for a week, j 

 It stains sections in from I to 5 minutes, and if too con-j 

 centrated may be diluted with freshly distilled water. 

 This solution does not readily deteriorate. Hamilton's 

 Hematoxylin. A rapid and strong nuclear stain 

 of the following composition: hematoxylin 12 gm. 

 ammonium-alum 50 gm., glycerin 65 c.c, distill© 

 water 130 c.c, carbolic acid 5 c.c. Heat the mix 

 ture to boiling before adding the carbolic acid, and 

 expose to sunlight for I month. Haug's Hema- 

 toxylin. A nuclear stain, giving good results wit! 

 nervous tissue. Dissolve I gm. of hematoxylin in K 

 c.c. of absolute alcohol, and add to 200 c.c. of an . 

 solution of aluminum acetate. Let the solution ripei 

 until the violet-black hue becomes brown-black. Aftei 

 staining, treat the sections with acid alcohol, and was! 

 in water until they appear blue. Heidenhain's Hema' 

 toxylin. Place the object to be stained in a 33 pe 

 cent, solution of hematoxylin in distilled water for fron 

 1 2 to 24 hours, and then for the same length of time in . , 

 0.5 percent, solution of neutral potassium chromate.am 

 wash out the excess of this with water, treating furthe ; 

 as desired. The stain is clear, from black to 

 color, rich in detail, and adapted to staining in mass 

 and has. the advantage that objects can be decolorized t< 

 any extent by prolonging the washing in the chromat" 

 If instead of this a I per cent, alum solution be used 

 the stain will be blue. Apathy's Modification. Maki 

 a 1 per cent, solution of hematoxylin in 70 01 

 cent, alcohol. After staining, wash out in a 1 1 

 solution of potassium bichromate i n from 70 to 80 1 

 alcohol. The mixture should be made freshly and k< 

 in the dark while using, as should also the objei 

 being washed, and they should be finished by washi 

 in several changes of 70 per cent, alcohol. Iron Hem 

 atoxylin. Sections are treated with a weak 

 solution of ferric acetate, washed in water, and 

 in 0.5 per cent, aqueous solution of hematoxylin, 

 blue-black or black-brown stain is obtained. This p it> 



