ELECTRON MICROSCOPY 



86 



ELEMENTARY BODIES 



times. By certain modifications the 

 magnification can be increased con- 

 siderably. It is feasible with this in- 

 strument to obtain photographs that 

 record the precise localization of cal- 

 cium and magnesium salts in tissues. 

 Scott and Packer {ibid, 31-45) showed 

 that the calcium and magnesium of 

 skeletal muscle was confined almost en- 

 tirely to the muscle fibers themselves, 

 and that in contracted fibers a great 

 concentration of magnesium appeared 

 in the contraction nodes. 



Tissues to be studied with this tech- 

 nique must be preserved in a manner 

 tliat permits no redistribution of min- 

 erals. The satisfactory method is that 

 of Altmann-Gersh. 



Electron microscopic technique sup- 

 plements histospectrography by pre- 

 cisely locating certain elements within 

 tissues and is very useful in conjunction 

 with the technique of microincineration 

 as a means of identifying certain com- 

 ponents of the ash seen in sections. 



Electrophoresis, Most particles suspended 

 in water carry electricity. If placed 

 in an electric field those possessed of 

 positive charge move toward the cath- 

 ode and those with a negative charge 

 toward the anode. Obviously there- 

 fore the nature of the charge and the 

 speed of movement can be determined 

 by microscopic study of particles sus- 

 pended in fluid in what is known as a 

 micro-electrophoresis cell. Types of 

 cell and precautions to be observed in 

 their use are described by Moore, D. H. 

 and Abramson, H. A. Glasser's Medical 

 Physics, 403-407. Their account of the 

 "moving boundary" method of electro- 

 phoresis and of the Tiselius apparatus 

 is clear and to the point. This latter 

 method, in contrast with the micro- 

 scopic one, affords a technique of great 

 accuracy and sensitivity for separating 

 concentrations and purifying submicro- 

 scopic components in blood serum and 

 other complete liquids. 



Eleidin (G. elaia, oil) gives to the stratum 

 lucidum its clear, glassy appearance. 

 It may be a dissociation product of 

 keratohyalin. There has been no great 

 improvement on the specificity of the 

 older methods. Mallory (p. 260) gives 

 the method of Buzzi (1889), first cau- 

 tioning that fixation must be in formalin, 

 Orth's or Bouin's fluid. Stain frozen 

 sections of 10% formalin fixed tissue in 

 sat. aq. picric acid (approximately 

 1.2%) 5 min. Rinse in aq. dest. and 

 counterstain for 1 min. in 1% aq. nigro- 

 sin. Wash in water and then in 95% 

 ale. (Skip absolute) Clear in ter- 

 pineol or origanum oil. Mount in bal- 

 sam: keratin, bright yellow; eleidin, 



blue black. Ranvier's Picro-Carmine 

 gives a fine red staining of eleidin. See 

 finger Nails. 

 Elementary Bodies are the smallest particles 

 of viruses. Those of certain viruses are 

 large enough for direct microscopic 

 examination in suitably stained prepara- 

 tions Vv'hich usually show also the larger 

 Inclusion Bodies if these are present. 

 Various methods designed for Rickettsia 

 are usually satisfactory. Many special 

 techniques have been proposed of which 

 2 follow : 



1. Methyl violet or Victoria blue 

 for smears (Gutstein, M., J. Path. & 

 Bact., 1937, 45, 313-314). Dry smears 

 on perfectly clean slides in air or incu- 

 bator. If necessary remove excess 

 protein by rinsing in physiological saline 

 solution followed by aq. dest. Fix in 

 methyl alcohol 1 hr. Stain in either of 

 2 ways: (1) Place slide in Petri dish. 

 Mix equal parts 1% aq. methyl violet 

 and 2% aq. NaHCOs. Filter imme- 

 diately onto the slide, cover dish and 

 incubate at 37 °C. 20-30 min. Rinse 

 in aq. dest., dry and mount in cedar oil 

 or liquid paraffin. Elementary bodies 

 light violet. (2) Same except filter 

 onto slide equal parts (a) Victoria blue 

 411 1 gm., ale. (abs.) 10 cc. and aq. dest. 

 90 cc. and (b) 0.02% aq. KOH and leave 

 at room temperature over night. Ele- 

 mentary bodies of vaccinia and other 

 viruses dark blue. 



2. Methyl blue acid fuchsin for sec- 

 tions (Nicolau, S. and Kopciowska, L., 

 C. r. Acad. d. Sci., 1937, 204, 1276-1278). 

 Fix in alcoholic Bouin's fluid. Stain 

 4-5 micron paraffin sections 30-60 min. 

 in: methyl blue (Grlibler) 1.5 gm., aq. 

 dest. 65 cc, methyl alcohol 35 cc, glyc- 

 erin 5 cc, 3% aq. oxalic acid 5 cc. 

 Wash well in aq. dest. and change to 

 absolute alcohol. Stain 20 min. in: 

 acid fuchsin 1.5 gm., aq. dest. 100 cc, 

 3% aq. oxalic acid 2 cc. Wash directly 

 in absolute alcohol and mount in the 

 usual way. Small particles in cells 

 associated with following viruses : 

 herpes, Borna, Zoster, rabies and 

 pseudo-rabies are stained bright red. 



A summary of methods for demon- 

 strating elementary bodies is given by 

 Seiffert, G., Virus Diseases in Man, Ani- 

 mal and Plant. New York : Philosophi- 

 cal Library, Inc., 1944, 332 pp. Under 

 favorable conditions some kinds of 

 of elementary bodies are visible at high 

 magnification unstained by both direct 

 and dark field illumination. Supra- 

 vital stains such as brilliant cresyl blue, 

 neutral red, methylene blue and azur it 

 are recommended. Before staining 

 smears, fixed in a variety of ways, pre- 

 treatment with 2.5% aq. potassium per- 



