VITAL STAINING 



262 



VITAMINS 



tribution of the dye in the tissues. 

 The skin, Icidneys, adrenals, liver, 

 spleen and bone marrow will be found 

 quite deeply colored while the nervous 

 system has escaped. The heaviest 

 accumulation will be in the peritoneal 

 cavity near the sites of injection and in 

 the loose connective tissue everywhere. 

 Examination of fresh mounts in physio- 

 logical salt solution will reveal that the 

 d}^e is concentrated within (1) the 

 epithelial cells of the convoluted tubules 

 of the kidney, of the adrenal and choroid 

 plexus; (2) certain cells of the ovary 

 and testicle; (3) the macrophages of 

 loose connective tissue and especially 

 of the spleen, liver, bone marrow, ad- 

 renals and lymph nodes — fibroblasts 

 are colored less deeply; and (4) the 

 "specific endothelia" of the five organs 

 mentioned. If permanent preparations 

 are desired fix in 10% formalin and im- 

 bed in paraffin. 



Vital staining in the narrow sense is 

 used for many purposes. (1) To iden- 

 tify phagocytic cells of the reticulo- 

 endothelial system and to see how they 

 behave in normal and pathological 

 conditions. (2) To locate injured cells 

 because some cells that do not ordi- 

 narily stain take up the dye when 

 injured. (3) To influence the activity 

 of R. E. cells by blocking them with 

 particulate matter. This has not been 

 very successful. See R. E. Blockade 

 (Victor, J., Van Buren, J. R. and 

 Smith, H. P., J. Exper. Med., 1930, 51, 

 531-548). (4) To measure the absorp- 

 tion by membranes of particulate matter 

 (Wislocki, G. B., Anat. Rec, 1921, 21, 

 29-33). (5) To distinguish between 

 malignant and non -malignant cells (Lud- 

 ford, R. J., Arch. f. exp. Zellf., 1933, 

 14, 42-55). (6) To determine pH of 

 different organs and tissues by injec- 

 tion with phthalein indicators (Rous, 

 P., J. Exper. Med., 1925, 41, 739-759). 

 (7) To identify calcium salts laid down 

 (Alizarin Red S and Madder). See 

 method for Reticulo-endothelial system. 



It is sometimes very worthwhile to 

 inject simultaneously three materials, 

 for example Higgins' Ink intravenously, 

 trypan blue or Niagara blue intraperi- 

 toneally, and lithium carmine intra- 

 pleurally (Foot, McClung, p. 116). 

 An interesting experiment is to feed 

 Sudan III or Scharlach (scarlet = 

 Sudan IV) colored lipids. IVIake solu- 

 tion in olive oil (about 20%). Intro- 

 duce by stomach tube into a cat. There 

 is slight staining of fatty tissue within 

 24 hrs. and maximum in 3-7 days 

 (Hadjioloff, A., Bull. d'Hist. Appl., 

 1938, 15, 81-98). Try also inducing 

 cat to drink large amount of milk or 



cream colored with Sudan III or Sudan 

 black, see colored illustrations of Gage 

 and Fish (S. H. and P. A., Am. J. Anat., 

 1924-25, 34, 1-81). History of vital 

 staining (Conn, H. J. and Cunningham, 

 R. S., Stain Techn., 1932, 7, 81-90, 

 115-119). See Chorioallantoic Mem- 

 brane, Carmine, Iridjgo-Carmine, 

 Manganese Dioxide, Higgins' Ink, 

 Protargol (silver). Lampblack, Leuco- 

 Dyes, Nuclei, Titanium Dioxide, Tho- 

 rium Dioxide, Copper, Platinum, Iron, 

 Mercury, Lymphatic Vessels. 

 Vitamins. (Revised by C. Carruthers, 

 Barnard Free Skin & Cancer Hospital, 

 St. Louis, May 29, 1946.) Some vita- 

 mins are susceptible of microscopic 

 localization. Deficiencies in most of 

 them leave structural imprints in the 

 tissues. A list may therefore be useful. 

 Up-to-date information is usually given 

 in Annual Review of Biochemistry. 

 See papers by P. Gj^orgy and R. A. 

 Morton in the re\dew for 1942, 11, 309- 

 364 and 365-390. A useful background 

 is provided by Sherman, H. C, Chemis- 

 try of Food and Nutrition, New York: 

 Macmillan, 1941, 611 pp. For a sum- 

 mary of tissue changes in vitamin 

 deficiencies see Wolbach, S. B. and 

 Bessey, O. A., Physiol. Rev., 1942, 22, 

 233-289. 

 A. Growth promoting, anti -infective 

 and anti-xerophthalmic vitamin. 

 C20H29OH, mol. wt. 286.4. The term 

 vitamin A is also applied to its provita- 

 mins: alpha-, beta- and gamma, caro- 

 tene and cryptoxanthin. There are 

 two tests for this vitamin. (1) The 

 antimony trichloride test is the basis of 

 the Carr-Price reaction, which see, as 

 applied to mitochondria of hepatic cells. 

 When the mitochondrial fraction is 

 separated and collected by Centrifuga- 

 tion the vitamin A can be easily 

 measured in it as the Goerner's have 

 done in their several investigations 

 (A. and M. M., J. Biol. Chem., 1937-38, 

 122, 529-538; ibid, 1939, 128, 559-565). 

 This test also has been employed for 

 vitamin A in serum the colors being 

 checked against alizarin solutions 

 (Parker, R. C, Methods of Tissue 

 Culture, New York: Hoeber, 1938, 292 

 pp.). According to Joyet-Lavergne, P., 

 C. rend. Acad. d. Sci., 1935, 201, 1219- 

 1221, vitamin A can be demonstrated 

 in the red blood cells of rays (marine 

 fish) by the antimony trichloride test. 

 (2) Green fluorescence. Much work has 

 been done on the identification of vita- 

 min A within cells bj'' its characteristic 

 but short lived fluorescence in ultra- 

 violet light. Popper, H., Proc. Soc. 

 Exp. Biol. & Med., 1940, 43, 133-136, 

 234-236 advises fixation of liver in 10% 



