SPIRIT BLUE 



229 



SPREADING FACTORS 



Spirit Blue (CI, 689)— anilin blue alcohol 

 soluble, gentian blue 6B, light, Lyon 

 and Paris blues — A mixture of di- and 

 tri -phenyl rosanilin chlorides. Conn 

 (p. 133) reports that it is a good stain 

 for growing nerve fibers. 



Spirit Indulin, see Indulin, spirit soluble. 



Spirit Nigrosin R, see Indulin, spirit soluble. 



Spirochaetales. The organisms of this 

 group often require special methods for 

 demonstration; but within the gastric 

 glands of humans (Doenges, J. L., Arch. 

 Path., 1939, 27, 469-477) dogs, cats, rats 

 and Macacus rhesus monkeys (Cowdry, 

 E. V. and Scott, G. H., Arch. Path., 

 1936, 22, 1-23) they can frequently be 

 seen in ordinary hematoxylin and eosin 

 preparations. Preparations of these be- 

 nign organisms are therefore easily 

 made and useful as showing intracellular 

 forms within parietal cells. For special 

 techniques see Treponema Pallida. 



Spleen. Fixatives penetrate the spleen 

 poorly on account of the large amount 

 of blood in it. Consequently it is desir- 

 able to fix only thin slices of it, say 3-4 

 mm. thick. If the spleen is particularly 

 soft to begin with the slices will not hold 

 their shape and it may be necessary to 

 cut parallel to the surface and include 

 the capsule as a support. Direct ob- 

 servation of splenic venous sinuses 

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

 64, 499-524; 65, 23-50; IVIacKenzie, D. 

 W., Whipple, A. O. and Wintersteiner, 

 M. P., Am. J. Anat., 1941, 68, 397-456). 

 Transplants into omentum (Holyoke, 



E. A., Am. J. Anat., 1940, 66, 87-132). 

 For vascular injections of Malpighian 

 bodies, see Nisimaru, Y. and Staggerda 



F. R., J. Physiol., 1932, 74, 327-337. 

 See Kurlof Bodies. 



Spodogramme, term used by French his- 

 tologists for the mineral skeleton of 

 tissue revealed by Microincineration. 



Spore Stain, a modification of Dorner's. 

 Make thin film on slide. Cover with 

 blotting paper and add freshly filtered 

 Ziehl's carbol fuchsin. Steam 5-10 min. 

 on hot plate, the blotting paper being 

 moistened with the fuchsin. Decolor- 

 ize instantaneously with 95% alcohol 

 and wash in water. Add drop of sat. 

 aq. nigrosine and spread thinly. Dry 

 quickly and examine. Spores red, other 

 parts of bacilli almost colorless against 

 dark background. Said to be simpler, 

 quicker than the unmodified Dorner's 

 method. It is recommended for Bacil- 

 lus megatherium, B. niger, B. cereus, 

 B. mycoides and some cultures of B. 

 subiilis (Snyder, M. A., Stain Techn., 

 1934, 9, 71-72). 



Stain heat fixed film with carbol- 

 fuchsin (see Acid Fast Bacilli). Rinse 

 quickly and differentiate in 95% alcohol. 

 Wash in hot tap water and again rinse 

 in alcohol. Counterstain for 2-5 min. 



with Loeffer's methylene blue. In case 

 of thick films pour off and add more 

 blue. Rinse in tap water and blot dry 

 (S. Bayne-Jones in Simmons and Gentz- 

 kow, p. 386). 



A modification of Schaeffer's spore 

 stain. Support a small metal tray over 

 asbestos centered wire gauze. Add 

 water and heat to steaming. Slides with 

 ends resting on either side of the tray 

 should have droplets of water condense 

 on their under surface. Flood properly 

 fi.xed smear on slide with 5% aq. mala- 

 chite green and leave in this way on 

 steam bath 1 min. Drop in cold water, 

 rinse thoroughly and while wet add 0.5% 

 aq. safranin 30 sec. Rinse again in cold 

 water. Spores, green; vegetative cells, 

 red (Ashby, G. K., Science, 1938, 87, 

 443). 

 Spreading Factors. The recognition of 

 these factors constitutes a major ad- 

 vance in biology and medicine. In this 

 Duran Reynals and his associates have 

 taken the lead. Those seeking informa- 

 tion as to techniques for the investiga- 

 tion of spreading factors should begin 

 with a detailed statement by Duran 

 Reynals, F., Bact. Rev., 1942, 6, 197- 

 252, on which the following account is 

 mainly based. 



Included under this heading are "sub- 

 stances present in animal tissues which 

 have the common property of increas- 

 ing the permeability of connective 

 tissue." By so doing they promote 

 spread through connective tissue of a 

 wide variety of substances, viruses and 

 microorganisms. Duran Reynals di- 

 vides them tentatively into 3 groups: 



1. "Factors with spreading power in 

 vivo and an enzymatic effect on hyal- 

 uronic acid in vitro as shown by reduc- 

 tion of viscosity and by hydrolysis," in 

 extracts of testicle, spleen, skin, etc. 



2. " Factors showing spreading power, 

 but no enzymatic activity in vitro, on 

 hyaluronic acid" in other tissues and in 

 some bacteria. 



3. Ascorbic acid, some other reducing 

 substances, possibly azoproteins. 



The factor acting on Hyaluronic Acid 

 is the enzyme hyaluronidase. The sub- 

 stance now known as hyaluronic acid 

 was demonstrated in the ground sub- 

 stance of connective tissue by staining 

 with toluidine blue by Bensley, S. H., 

 Anat. Rec, 1934, 60, 93-108. The same 

 investigator studied the consistency of 

 the ground substance by the ingenious 

 device of first adapting paramoecia to 

 tissue fluid and then of observing that 

 their movements in the intercellular 

 and interfibrous spaces of connective 

 tissue are restrained by invisible mate- 

 rial. See Loose Connective Tissue. A 

 specific microchemical test is greatly 



