[CITATION][C] The reticulum of lymph nodes in mice studied with the electron microscope

SL Clark Jr - American Journal of Anatomy, 1962 - Wiley Online Library
SL Clark Jr
American Journal of Anatomy, 1962Wiley Online Library
MATERIALS AND METHODS Mesenteric and occasionally other lymph nodes were
obtained from 11 mice of various strains, fixed one or two hours at room temperature in 1%
osmium tetroxide, rinsed in water, dehydrated by serial dilutions in ethanol, and imbedded in
a mixture of methyl and N-butyl methacrylate, polymerized with heat and benzoyl peroxide.
For fixation, the osmium tetroxide was dissolved in three different buffers: the veronal-
acetate mixture of Palade ('52) to which 3.5% sucrose had been added, Dalton's potassium …
MATERIALS AND METHODS
Mesenteric and occasionally other lymph nodes were obtained from 11 mice of various strains, fixed one or two hours at room temperature in 1% osmium tetroxide, rinsed in water, dehydrated by serial dilutions in ethanol, and imbedded in a mixture of methyl and N-butyl methacrylate, polymerized with heat and benzoyl peroxide. For fixation, the osmium tetroxide was dissolved in three different buffers: the veronal-acetate mixture of Palade ('52) to which 3.5% sucrose had been added, Dalton's potassium dichromate solution ('55), and a balanced salt solution designed for tissue culture by White ('54), but adapted as a diluent for osmium tetroxide by Dr. Walter C. Bauer (personal communication,'60). The solution contained all of the the constituents specified by White except ferric nitrate, and its pH was 7.4, In some instances methyl cellulose was added to a concentration of 0.5% and sometimes the fixative was cooled to 0 C.
Thin sections were cut with glass knives in a Porter-Blum microtome (Servall) and examined in an RCA EMU 2E electron microscope. In order to enhance the contrast of connective tissue fibers, some sections were stained on the supporting grid by floating for one to three hours on a saturated solution of uranvl acetate and rinsing in distilled water. Tne destructive effects of the electron beam were mitigated, in some instances, by covering stained sections with a film of collodion
Wiley Online Library