RICKETTSIA ORIENTALIS 



308 



ROULEAUX FORMATION 



95% alcohol. Counterstain with 0.25% 

 aq. basic fuchsin for 30 min. Decolor- 

 ize quickly (say 3 sec.) in 0.5% aq. 

 citric acid. Differentiate rapidly in 

 abs. ale. clear in xylol and mount in 

 dammar. Rickettsiae, deep red; sur- 

 rounding tissue, partly red. Back- 

 ground can be made bluer by washing 

 lightly in aq. dest. after the citric acid 

 treatment and by staining again with 

 methylene blue, before differentiation 

 in 95% alcohol dehydration, clearing 

 and mounting as advised by Plotz. 

 Plotz gives details of use of Michia- 

 vello's stain in demonstration of Rick- 

 ettsiae in yolk sac cultures. 



A fuchsin and methyl violet combina- 

 tion is recommended for typhus fever 

 Rickettsiae by Nyka, W., J. Path. & 

 Bact., 1944, 56, 264. 



See cultivation of Rickettsiae in eggs 

 (Fitzpatrick, F. K., J. Lab. & Clin. 

 Med., 1946, 31, 45-55), Typhus Fever 

 rickettsiae, and Rickettsia orientalis. 

 A convenient list of pathogenic Rick- 

 ettsia is provided by Pinkerton, H., 

 Bact. Rev., 1942, 6, 37-78. 



Rickettsia orientalis. Rapid method for 

 staining in smears by Clancy, C. F. and 

 Wolfe, D. M., Science, 1945, 102, 483. 

 Air dry smears of infected yolk sac 

 membranes, or of other tissues, and fix 

 by heat. Flood slide with xylol, dry 

 in air current, immerse in 1:5,000 

 methylene blue and basic fuchsin in aq. 

 dest. for 5 min. Wash, dry and ex- 

 amine. Organisms blue, background 

 pinkish purple. Grams should be di- 

 luted from 1% stock solutions on the 

 day used. 



Ringer solution. NaCl, 0.85 gm.; KCI, 

 0.025 gm.; CaCU, 0.03 gm.; aq. dest., 

 100 cc. Lee (p. 731) advises for am- 

 phibians same except that NaCl is 0.65 

 gm. and NaHCOs, 0.02 gm. is added to 

 make pH about 7.0-7.4. If NaHCOs is 

 present it must not be sterilized by 

 heat. 



Ringer-Locke solution. NaCl, 0.85 gm.; 

 KCI, 0.042 gm.; CaCh, 0.025 gm.; 

 NaHCOs, 0.02 gm.;aq. dest., 100 cc. for 

 cold blooded animals. Lee (p. 73) ad- 

 vises same except that NaCl is 0.65 gm. 

 Should be freshly made. Owing to 

 presence of NaHCOa must not be steri- 

 lized by heat. 



Rivanol, a dye sometimes employed in 

 fluorescence microscopy. 



Roberts, see Paper Chromatography. 



Rocky Mountain Spotted Fever, see Rick- 

 ettsia. 



Roller Tube Cultures. Control of pH in, see 

 paper by Paff, G. H., Proc. Soc. Exp. 

 Biol. & Med., 1946, 62, 184-187. See 

 Tissue Culture. 



Romanowsky Stains contain polychrome 



methylene blue eosinates. Those of 

 Wright, Leishman and Wilson are well- 

 known. The Romanowsky effect is 

 the lavender-red coloration by them of 

 the nuclei of lymphocytes, monocytes, 

 protozoa and other materials. Acetone 

 solvents for Romanowsky stains (Kings- 

 ley, D. M., J. Lab. & Clin. Med., 

 1936-37, 22, 524-531). Polychroming 

 process (i6 id, 736-752). Dyes for (ibid, 

 1264-1273). Large bibliographies. 



Romieu Reaction for proteins. Fix in 

 formalin, in alcohol or in Bouin's fluid. 

 Make rather thick sections in paraffin 

 or preferably in celloidin. Cover sec- 

 tion with a drop of syrupy phosphoric 

 acid. After few minutes in oven at 

 56°C. examine directly. A red or 

 violet color develops in location of pro- 

 teins. According to Blauchetiere and 

 Romieu (A. and B., C. Rend. Soc. de 

 Biol., 1931, 107, 1127) it is due to the 

 tryptophane grouping. See Lison, p. 

 129. 



Rongalite White, said to stain normal but 

 not cancerous cells (Roskin, G., Bull. 

 d'Hist. appl., 1938, 15, 20-23). 



Rosanilin (Magenta I) is triamino-tolyl- 

 diphenyl-methane chloride, a compo- 

 nent of most Basic Fuchsins. Rosan- 

 ilin with methylene blue for Negri 

 bodies (Schleifstein, J., Am. J. Pub. 

 Health., 1937, 27, 1283-1285). 



Rosazine, see Azocarmine G. 



Rose Bengal (CI, 779). A xanthene dye of 

 fine color used for several purposes 

 including the staining of Soil Bacteria 

 by Conn (p. 157). Make suspension 

 of soil in 9 times its weight of 0.015% 

 aq. gelatin. Spread drop on clean slide 

 and dry over boiling water bath. Cover, 

 while still on bath for 1 min., with rose 

 bengal 1 gm.; CaClj, 0.01 gm.; 5% aq. 

 phenol, 100 cc. Wash quickly in water. 

 Dry and examine. Used to demon- 

 strate injured liver cells by vital stain- 

 ing (Williams, W. U., Yale J. Biol, and 

 Med., 1950, 23, 177-183). See Eosins. 



Rosenthal, see Fluorescence, Microscopy, 

 Radioactive Isotopes, Electron Micros- 

 copy. 



Rosin U.S. P. XI (Colophony, yellow resin, 

 abietic anhydride) used in Grieves' 

 method for undecalcified dental tissues 

 and bone. 



Rosinduline GXF, see Azocarmine G. 



Rosophenine lOB, see Thiazine Red R. 



Rouget Cells, see Pericapillary cells. 



Rouleaux Formation of red cells. They 

 are stacked like coins, outlines distinct, 

 usually break up on shaking, diminished 

 on washing with saline solution; 

 whereas agglutinated reds stick to- 

 gether irregularly, outlines indistinct, 

 usually become more compact on shak- 

 ing, not affected by washing. 



