TRANSPARENT CHAMBER 

 TECHNIQUES 



356 



TREPONEMA PALLIDUM 



the preparation of a lucite calvarium, 

 in which the convex portion of the skull 

 of a monkey is permanently replaced 

 by a transparent lucite plate, thus ex- 

 posing the surface of both cerebral 

 hemispheres. These preparations have 

 been used in studies of superficial 

 cerebral vessels in head injury, drug 

 administration, and oxygen poisoning. 



Several attempts have been made to 

 install permanent type chambers for 

 the study of internal organs. Zintel 

 (Anat. Rec, 1936, 66, 437-447) intro- 

 duced one of the earliest of these for 

 exteriorizing a loop of intestine and its 

 mesentery. A chamber for the 

 pancreas of the mouse has been de- 

 scribed (Flory, C. M. and Thai, A., 

 Anat. Rec, 1947, 97, 33-40). Estable 

 (Proc. Soc. Exp. Biol, and Med., 1948, 

 67, 445-447) has described a technique 

 for biomicroscopic study of the ovary 

 and the Fallopian tube in rabbits. 

 The ovary is transferred to a subcu- 

 taneous space, retaining intact its 

 vascular and nerve supply. A trans- 

 parent capsule is applied for protection 

 during repeated microscopical examina- 

 tion. 



Adaptation of the transparent cham- 

 ber technique to mice (Algire, G. H., 

 J. Nat. Cancer Inst., 1943, 4, 1-11); 

 Algire, G. H. and Legallais, F. Y., J. 

 Nat. Cancer Inst., 1949, 10, 225-243) 

 makes the procedure especially useful 

 in the many phases of cancer research 

 involving inbred strains of mice. In 

 this modification tantalum sutures are 

 used to support a dorsal fold of two 

 skin layers between two plastic (vinyl- 

 ite) splints. One skin layer is incised 

 and retracted to allow the introduction 

 of a lucite ring and attached coverslip 

 which is in contact with the inner sur- 

 face of the second layer of skin. Both 

 the lucite ring and the supporting 

 splints are held in position by tantalum 

 bolts. This procedure makes acces- 

 sible to study by transmitted light a 

 layer of tissue approximately 500 micra 

 thick, consisting of peripheral nerves, 

 striated muscle, peripheral blood ves- 

 sels and lymphatics, subcutaneous 

 connective tissue and fat, hair follicles, 

 and epidermis. 



Implantation of normal and neoplas- 

 tic cells, of embryonic organs, or of 

 carcinogenic chemicals is readily per- 

 formed during the operation for in- 

 sertion of the chamber. Experimental 

 studies of the tissue may be under- 

 taken immediately after the operation, 

 and carried on for from 30 to 60 daj^s. 

 Response of the host and the implanted 

 tissues to physical and chemical agents 

 may be studied in terms of cellular and 



circulatory reactions, including quanti- 

 tative measurements of blood pressure, 

 dye diffusion, and arterial oxygen 

 saturation. Microscopic resolution is 

 sufficiently good to observe cross-stria- 

 tions in muscle, platelets in circulating 

 blood, and cytoplasmic detail in cells 

 adhering to the coverslip. Additional 

 resolution of cytologic detail has been 

 obtained through use of a round-table 

 design. The access-type consists of a 

 thick (1 mm.) coverslip of lucite 

 through which a 1 mm. diameter hole 

 provides for local application of chem- 

 icals at any time after introduction 

 of the chamber. The operative pro- 

 cedure requires less than one hour and 

 two workers can readily handle 10 to 

 15 animals in daily observations, 

 measurements, and photographic rec- 

 ords. 



The dynamic, functional aspects of 

 the transparent-chamber approach in- 

 dicates future increased development 

 and correlations with other methods 

 of biological research. 



Trematodes. Make up stain by mixing 

 1 gm. of dried residue on filter paper 

 from Schneider's aceto-carmine with 

 10 gm. ammonia alum in 200 cc. aq. 

 dest. with aid of heat. When dissolved, 

 cool, filter and to filtrate add crystal 

 of thymol. After fixation bring worms 

 to water or to 20% alcohol. Stain 12-36 

 hrs. depending on size. Remove to 

 water 2 changes. Dehydrate through 

 20, 35, and 50 to 70% alcohol. Place 

 few crystals potassium chlorate in small 

 glass covered dish; add few drops cone. 

 HCl. When chlorine is given off fill 

 dish with 70% alcohol. If deeply 

 stained differentiate in this chlorinated 

 alcohol. If not or the specimens are 

 small ones add it to the alcohol covering 

 them and agitate. When sufficiently 

 destained remove to fresh 80% alcohol. 

 Dehydrate in alcohol. Add cedar wood 

 oil to the absolute until mixture is one 

 half oil. Clear in cedar oil and mount 

 in balsam (Gower, W. Carl, Stain 

 Techn., 1939, 14,31-32). 



Treponema Pallidum. The organisms can 

 best be seen in the primary lesions by 

 Darkfield examination. The same 

 method is useful for skin and lymph 

 nodes in the secondary stage but for 

 the tertiary lesions in deep lying tissues 

 sections are desirable supplemented 

 by smears. A negative finding is com- 

 forting but does not necessarily signify 

 absence of parasites unless confirmed 

 serologically. 



1. Low surface tension stain for 

 smears (Haire, R. D., J. Lab. & Clin. 

 Med., 1938, 23, 1215-1216). Mix 1 gm. 

 Gentian violet (or crystal violet) in 



