STAINS, ETC. 



1386 STAINING OF CONNECTIVE TISSUES. 



injected preparations are thrown into strong alcohol or 

 chromic acid, which sets the mass. Hoyer's Oil- 

 color Masses. Mix with 30 parts of lavender, fen- 

 nel, thyme, or rosemary oil, 5 parts of artists' Berlin 

 blue oil-color, rubbed up with 5 parts of thickened 

 linseed-oil. Let the mixture stand 24 hours in a closed 

 vessel ; then decant. Shake before using. This is 

 useful for injecting the vessels of the spleen and other 

 structures difficult of injection. Hoyer's Shellac 

 Mass. Place in a wide-necked flask a quantity of good 

 shellac, with enough 80 per cent, alcohol to cover it. 

 After 24 hours, warm it on a water-bath, to complete 

 the solution ; cool, dilute with alcohol to a thin syrupy 

 consistence, and strain through thick muslin. Color 

 the solution with anilins in filtered concentrated alco- 

 holic solution. Cinnabar may be used for corrosion- 

 preparations. Berlin blue and yellow arsenic sulpbii 

 are useful ; both yield a green color. The pigmerts 

 should be rubbed to fine powder with water, and 

 alcohol added. When the mixture has settled, pour 

 off the dilute and add strong alcohol. By shaking 

 the flask the coarser particles settle ; pour off the fluid 

 containing the finer ones, add it to the shellac solution, 

 and strain through muslin. Hydrochloric acid does 

 not attack this solution ; hence it is useful for corrosion- 

 preparations. Joseph's White-of-egg Mass. Take 

 filtered white-of-egg and dilute it with I to 5 per cent, 

 of carmin solution. This mass remains liquid when 

 cold. It coagulates when immersed in dilute nitric, 

 chromic, or osmic acid, is transparent, and indifferent 

 to reagents. It is useful for Invertebrates. Robin's 

 Gelatin Vehicles. I. Soak 1 part of " colle de 

 Paris ' ' gelatin in 10 parts of cold water ; heat 

 in a water-bath, and add 2 per cent, of chloral as a 

 preservative. 2. Dissolve in a water-bath 50 gm. of 

 "colle de Paris" gelatin in 300 gm. of water con- 

 taining a little arsenious acid ; add a few drops of car- 

 bolic acid and 150 gm. of glycerin. This does not 

 keep as well as the pure gelatin vehicle. Scheele's 

 Green Mass. a. Eighty c.c. of a saturated solution 

 of potassium arseniate and 5° c - c - of glycerin, b. 

 Forty c.c. of a saturated solution of copper sulphate 

 and 50 c.c. of glycerin. Combine the two solutions 

 with three volumes of the vehicle. 



STAINING OF CONNECTIVE AND OTHER 

 TISSUES. 

 Areolar Tissue. Inject hypodermatically into the sub- 

 cutaneous tissue of a dog or rabbit a I : 1000 solution 

 of silver nitrate. With a pair of curved scissors snip 

 off a little of the edematous tissue, and stain with 

 picrocarmin for from 10 to 12 hours in a moist cham- 

 ber. The fibrous and cellular elements are then brought 

 into view by treating with glycerin slightly acidulated 

 with formic acid. Bile-capillaries. Golgi's Method. 

 Fix small cubes of liver 3 to 4 days in a mixture of 4 

 parts of a 3 per cent, solution of potassium bichromate 

 and I part of a I per cent, osmic acid ; then place in a 

 0.75 per cent, solution of silver nitrate for two days, 

 wash in distilled water, and harden in alcohol. Sec- 

 tion and mount in balsam. The capillaries appear as 

 a black network on a yellow ground. Bone. 1. 

 Fle/nming's Method. Soak sections of decalcified 

 bone in water, and place in a drop of water on a glass 

 plate ; remove the excess of water with bibulous paper 

 and cover with another glass plate to prevent rolling ; 

 place the whole in a dish and cover with alcohol. In 

 half an hour the sections will be fixed and flat. Place 

 in absolute alcohol. To mount, wash in fresh alcohol, 

 then in ether ; place the sections on glass, cover with two 

 thicknesses of blotting-paper and a glass plate, and dry 

 for a day in the air or in an oven. Put a drop of melted 



balsam on a slide and another drop on a cover-glass ; 

 place the section on the slide, cover, put on a clip, 

 and warm. 2. White's Method. Suitable for osseous 

 or dental tissue. Sections ground moderately thin are 

 soaked in ether for 24 hours, then placed for 2 01 3 

 days in a thin solution of collodion stained with fuch- 

 sin, then hardened in alcohol, ground to the requisite 

 thinness between two plates of ground glass, with 

 water and pumice powder, and mounted, dry, in thick 

 balsam. The stained collodion is prepared by dissolv- 

 ing fuchsin in methylated spirit and adding the ether 

 and pyroxylin. 3. Vivante 's Method. Place very 

 small pieces of young bone for 8 days in Miiller's 

 fluid, then in the osmium- bichromate mixture, then in 

 silver solution. After impregnation decalcify for 20 

 days in von Ebner's fluid ; then wash in water, place in 

 a solution of sodium carbonate, and embed in paraffin. 

 Cartilage. 1. Ranvier ' s Method. Place sections of 

 fresh cartilage for 24 to 48 hours in a few c.c. of 

 Ranvier's purpurin solution (see Staining Reagents), 

 wash in water, and mount in glycerin. The nuclei are 

 stained, the matrix remaining almost colorless. 2. Rub 

 the cartilaginous end of the freshly excised femur of a 

 frog " 7 ith a stick of silver nitrate, and expose to sun- 

 light Section, and mount in Farrant's solution. The 

 matrix is stained brown, and the apparently empty 

 spaces contain the cells, which are too transparent to 

 be readily seen. Columnar Cells. Wash a piece 

 of the mucosa of the small intestine of a cat in dis- 

 tilled water, place for 10 minutes in 0.5 per cent, 

 silver-nitrate solution, and silver in the usual way. 

 Harden in alcohol, detach the epithelium, mount in 

 glycerin. A view is obtained of the free ends of the 

 cells with the cement-substance between them as "sil- 

 ver lines," and also of the open mouths of the goblet- 

 cells. Cornea. Klein ' s Method. Remove from a 

 living cornea, by brushing, the conjunctival epithelium, 

 and rub the corneal surface with a stick of silver 

 nitrate ; in half an hour detach the cornea, and 

 examine it in distilled water. Negative images of the 

 corneal cells are thus obtained. To obtain positive 

 images, treat according to Ranvier's gold chlorid 

 lemon-juice method (see Staining Reagents, Metallic 

 Stains). Rolletf s Method. Immerse a fresh cornea 1 

 in aqueous humor, place it in a moist chamber, and 

 pose to the action of iodin vapor; when brown, peel 

 off the epithelium and examine. A good method, the 

 result being almost equal to that of the gold method. 

 Corpuscles of Grandy. Remove the skin and pa- 

 pillre from the margins of the fresh beak of a duck. 

 put pieces into 50 per cent, formic acid for 20 minul 

 or until transparent; remove the corneous lave 

 epithelium, rinse in water, and treat with gold chlo 

 according to Pritchard's method (see Staining 

 Metallic Stains). The same method may Ix 

 the corpuscles of Herbst. Elastic Tissue. 1. Mar- 

 tin Mi's Method. Fix for 3 weeks in 2 p- 

 acid, wash, and stain 48 hours in 5 per cent 

 safranin solution (see Staining Reagents). 1 

 fibers appear of an intense black, the other 1 issue- show 

 ing the usual tints of safranin staining. 2. I'mui 

 Method. Dissolve 0.1 gm. of orcein (GrUbler) in 20 

 gm. of 95 per cent, alcohol and 5 gm. of water; (Us 

 solved gm. of strong hydrochloric acid in a like 1 

 ture of alcohol and water. Take a numl>ero! 

 glasses, and pour IO drops of the stain in each ; 

 5 drops of the acid mixture to the first glass, 6 to th< 

 next, and so on, increasing the proportion by one dfl 

 until all are acidulated. In each glass place 1 

 2 sections, and stain 12 hours. Examine in ;i d»] 

 of glycerin ; the elastic fibers appear a shiny brown 1 

 lighter ground. Epithelium. A'romaver 's Method. 



