MALARIA PLASMODIA 



190 



MALLORY-HEIDENHAIN STAIN 



1942, 57, 90-94) find that when the 

 pigment within the parasites (Plas- 

 modium Knowlesi) is extracted in 

 such a way as not to influence the 

 spectra of hemoglobin it can be identified 

 spectrophotometrically as ferrihemic 

 acid, or hematin, which does contain 

 iron. 

 Malaria Plasmodia. Technique of examina- 

 tion of process of "exflagellation" 

 (Anderson, Ch. W. and Cowdry, E. V., 

 Arch, de I'lnst. Pasteur de Tunis, 1928, 

 17, 46-72), of quantitative determina- 

 tions of gametocytes (Cowdry, E. V. 

 and Covell, W. P., Ibid., 147-456) and 

 of demonstrating neutral red granules 

 and Golgi apparatus (Cowdry, E. V. 

 and Scott, G. H., Ibid., 233-252). 



For staining the plasmodia in smears, 

 see Giemsa, Jenner, Marino, Nocht, 

 Plehn, Wilson and Wright's stains. A 

 simple method for staining plasmodia 

 in paraffin sections is described with 

 numerous illustrations by Tomlinson, 

 W. J. and Grocott, R. G., Am. J. Clin. 

 Path., 1944, 14, 316-326. The Barber 

 Eomp thick film method is strongly 

 recommended for surveys. 



Serlin, N. J. and Lissa, J. R., Am. J. 

 Clin. Path., 1942, 6, 8 advise the follow- 

 ing method when diagnosis depends on 

 finding gametocytes, or malarial pig- 

 ment, in peripheral blood. Completely 

 evaporate 1 cc. 1% aq. potassium oxa- 

 late in a 15 cc. centrifuge tube. Add 10 

 cc. venipuncture blood. Mix carefully 

 and centrifuge 30 min. at 2,500 R.P.M. 

 Pipette oil all but about J in. of super- 

 natant plasma. Smear on 2 slides by 

 wiping buffer layer with stick applicator 

 having non-absorbent cotton tip. 

 Stain by Wright's method. Study of 

 Giemsa stained smears by dark field is 

 suggested (Goosmann, C., J. Lab. & 

 Clin. Med., 1935-36, 21, 421-424). See 

 Protozoa. 



Taylor, D. J., Greenberg, J. and 

 Josephson, E. S. (J. Lab. Dis., 1951, 88, 

 158-162) describe a useful method for 

 the maintenance of intraerythrocytic 

 forms of Plasmodium gallinaceum in a 

 whole medium on vitro. 

 Mallory's Connective Tissue Stain. This 

 is name usually given to his anilin 

 blue-acid fuchsin-orange G stain. See 

 also his Phosphomolybdic and Phospho- 

 tungstic Acid Hematoxylin Stains. 

 (Mallory, p. 155). Fix in Zenker's 

 fluid. Imbed in paraffin or celloidin. 

 Remove mercury from sections with 

 iodine or 0.5% sodium hyposulphite. 

 Stain in 0.5% aq. acid fuchsin, 1-5 min. 

 Drain off stain and put in : anilin blue, 

 water soluble, 0.5 gm. ; orange G, 2 gm. ; 

 1% aq. phosphotungstic acid, 100 cc, 

 20 min. or longer. Rinse in 95% ale. 



2 or 3 changes until no more stain is 

 removed. Dehydrate in abs. ale, clear 

 in xylol, mount in neutral balsam. For 

 celloidin sections, reduce staining time 

 and pass from 95% ale. to terpineol and 

 mount in balsam. This is one of the 

 most beautiful of all stains and is very 

 widely used. Collagenic fibrils blue, 

 fibroglia, neuroglia and myoglia fibrils 

 red, elastic fibrils pink or yellow. In 

 McClung, p. 405, Mallory and Parker 

 advise 0.25% aq. acid fuchsin and 

 staining in the anilin blue mixture for 

 1-24 hrs. or for 1 hr. in paraffin oven at 

 60 °C. The modifications of this stain 

 are almost endless. 



Adaptation to formalin fixed material 

 is often desirable. Kernohan (J. W., 

 J. Tech. Meth., 1934, 13, 82-84) has 

 outlined the following method of doing 

 this by mordanting. Wash formalin 

 fixed tissue in running water or in 

 ammonia water for short time. Place 

 in Weigert's primary mordant — potas- 

 sium bichromate, 5 gm.; chromium 

 fluoride, 2 gm. and aq. dest. 100 cc. — 

 for 4 days and in his secondary mordant 

 — copper acetate, 5 gm.; chromium 

 fluoride, 2.5 gm.; acetic acid (36%), 

 5 cc; aq. dest., 100 cc. and formol, 

 10 cc. — for 2 days. Imbed in paraflin 

 in the usual way. 



Rexed, B., and Wohlfart, G., Zeit. 

 wiss. Mikr., 1939, 56, 212-215 suggest 

 control of pH of the acid fuchsin. It is 

 stated that fresh 0.1% acid fuchsin has 

 pH 4.49 and that increase in alkalinity 

 makes it defective. To prepare one at 

 pH 3.29 ± 0.01, which is recommended, 

 take acid fuchsin 1 gm.; N/10 HCl, 60 

 cc. ; aq. dest. 900 cc. ; Storensen's citrate 

 (citric acid crystals, 21 gm.; N/1 

 NaOH, 200 cc; + aq. dest. to make 

 1000 cc), 40 cc. Most tissues stain in 

 range pH 3-4, red blood cells alone at 

 pH5-7. 



In 1936, Mallory considered (Stain 

 Tech., 11, 101-102) the most important 

 modifications of his stain to be Heiden- 

 hain's Azocarmine (Azan), the Lee- 

 Brown and Masson Trichrome methods. 

 See Grossman's modification and Pitui- 

 tary for special adaptations. 

 Mallory-Heidenhain Rapid One-Step Stain 

 for Connective Tissue — Written by 

 Jane E. Cason, Dept. of Pathology, 

 Medical College of Alabama, Birming- 

 ham, Ala. January 27, 1951— Although 

 innumerable modifications of Mallory's 

 aniline blue collagen stain for tissue 

 sections prevail, this procedure appears 

 to be an improvement over the original 

 and subsequent modifications. 



The necessity for adjusting the in- 

 tensity of the aniline blue and the acid 

 fuchsin in routine staining suggested 



