VAN GIESON'S CONNECTIVE 



259 



VERHOEFF'S ELASTIC TISSUE 



95% ale, dehydrate, dear and mount. 

 Muscle yellow, collagcnic fibers red, 

 nuclei blue black. A brilliant stain. 

 But it fades quickly and is not so much 

 employed at present as Mallory's con- 

 nective tissue stain. See Buzaglo's 

 Method, Curtis' Substitute for Van 

 Gieson, Collagenic Fibers, Connective 

 System. 

 Van Wijhe's method for staining cartilage 

 in whole tissues with methylene blue. 

 See Cartilage. 



Vasa Vasorum. Injection with India ink 

 (Winternitz, M. C, Thomas, R. M. 

 and LeCompte, P. M., The Biology of 

 Arteriosclerosis. Springfield: Thomas, 

 1938, 142 pp.). Filter Higgins Engros- 

 sing ink through coarse filter paper and 

 dilute filtrate with 8 times volume of aq. 

 dest. Obtain pressure apparatus con- 

 sisting of 2 liter metal tank with top 

 and bottom outlets and air pressure 

 gauge. Connect upper outlet with 

 escape valve and high pressure air line 

 and the lower one with rubber tube and 

 cannulae. To inject vasa of coronary 

 arteries place fresh human heart un- 

 opened in 0.9% aq. sodium chloride 

 containing 0.1% sodium nitrite and a 

 little thymol for 24 hrs. at 3-4°C. Just 

 before injection warm heart to 37 °C., 

 tie cannulae in openings of coronary 

 arteries and clamp or ligate all openings 

 of heart except the aorta. By opening 

 and closing the escape valve the ink in 

 the tank is driven into the coronaries 

 by a pulsating pressure. During first 

 10 min. maintain the minimum pressure 

 at about 100 mm. of mercury with 

 maximum pressure of pulsations not 

 more than 200. Then increase slowly 

 so that during next 20 min. the mini- 

 mum pressures vary 500-800 mm. and 

 the maximum 800-1000. After injec- 

 tion put heart in 10% formalin for 24 

 hrs. Dissect out main coronaries. 

 Clear by Spalteholz Method for whole 

 mounts or imbed in paraffin section and 

 color by Masson's Trichrome stain. 

 The authors give special directions for 

 injecting the aorta and vessels of kid- 

 neys and amputated legs. Their illus- 

 trations afford useful guides to the 

 results expected. 



Vaseline in tissues can be distinguished 

 from the normal fats by the fact that 

 the former is colored clear violet and 

 the latter intense blue black by stain- 

 ing for 15 min. with Sudan Black B. 

 Terebenthine, turpineol and methyl 

 benzoate are colored blue black (Gerard, 

 P., Bull. d'Hist. Appl., 1935, 12, 92-93). 



Vegetative Intermitotics, see Cell Classifica- 

 tion. 



Veins, see Blood Vessels and a very fine 

 presentation by Franklin, K. J., A 



Monograph on Veins. Springfield: 

 Thomas, 1937, 410 pp. with hundreds 

 of references to techniques and results. 



Venous Sinuses, splenic, direct observa- 

 tion in vivo (Knisely, M. H. Anat. 

 Rec, 1936, 64, 499-524; 65, 23-50). See 

 Spleen. 



Venules. A grapliic demonstration of 

 venules in the ears of white mice can 

 be obtained by intravenous injection 

 of Chicago blue because this dye escapes 

 into the surrounding tissue fluid more 

 easily from venules than from capil- 

 laries (Smith, P. and Rous, P., J. Ex- 

 per. Med., 1931, 54, 499-514). 



Verhoeflf's Elastic Tissue Method (Ver- 

 hoeff, F. H., J. A. M. A., 1908, 50, 

 876-877). Gives good results after 

 fixation in Zenker's fluid, formalin 

 alone or after Weigert's mordant for 

 mj^elin sheaths or Marchi's fluid. It 

 is fairly satisfactory for tissues decalci- 

 fied with nitric acid. Mercury deposits 

 resulting from Zenker's fixation are 

 removed by the stiiin : Hematoxylin 

 crystals, 1 gm.; Abs. ale, 20 cc; Dis- 

 solve in test tube with aid of heat, 

 filter and add in order given: 10% aq. 

 ferric chloride, 8 cc; Cone. Lugol's 

 solution (iodine, 2; potassium iodide, 4; 

 water, 100), 8 cc. Stain sections in 

 above sol. 5 min. or more. Differenti- 

 ate in 2% aq. ferric chloride for a few 

 sec. until the connective tissue takes 

 the color of Lugol's solution. Keep 

 sections in motion during differentia- 

 tion. They can be examined at low 

 magnification in water and if over 

 differentiated can be restained at this 

 stage. Wash in water followed by 95% 

 ale. to remove the stain of Lugol's solu- 

 tion. Then leave in water 5 min. or 

 more. Counters tain in 0.2% water 

 sol. eosin in 80% alcohol. Dehydrate, 

 clear in origanum and mount in balsam. 

 Elastic tissue, black; fibroglia, myoglia, 

 neuroglia, mj'^elin and fibrin, pink. 

 Degenerated elastic tissue (elacin) 

 can be distinguished by less intensity 

 of staining and by diffuse outlines. 

 To differentially stain myelin sheaths 

 fair results are obtained after Zenker's 

 fixative or formalin followed by Alarchi's 

 fluid. For best results fix in formalin 

 4 days, or longer, and mordant in 

 Weigert's potassium bichromate and 

 chrome alum for 4 days. Again it is not 

 necessary before hand to remove mer- 

 curial precipitates. Place sections in 

 3% aq. potassium permanganate, 30 

 min. Wash in water and color for 30 

 min. in the hematoxylin stain described. 

 Wash in water and differentiate in 10% 

 aq. ferric chloride until the internal 

 elastic membranes of blood vessels are 



