US EPA

6607168 

Journal Article 

Low-temperature embedding in water-miscible meth-acrylates after treatment with antifreezes 

Cope, GH 

1968 

88 

(2) 

235-257 

BCI:BCI19684900124798 

Mouse tissues were infiltrated at 0[degree]C with increasing concentrations of buffered aqueous solutions of non-electrolyte antifreezes. Up to 50% vv of dimethyl sulphoxide (DMSO), or mixtures of equal parts of DMSO and either glycerol or ethylene glycol were used. The specimens were then cooled slowly to -20[degree]C, (at a rate of less than 1[degree]C/min. overall) and the tissues were then fixed, dehydrated, and embedded in a liquid state at -20[degree]C. Tissues were fixed with 5% glutaraldehyde in 50% antifreeze solution for 6-8 hr. Tissues were then dehydrated in 2-hydroxyethyl methacrylate and embedded in mixtures containing 70% 2-hydroxyethyl methacrylate and 30% of either n-butyl methacrylate or styrene. Embedded tissues were polymerized with ultraviolet (UV) light at -10[degree]C. A modification in which tissues were substituted with a mixture of ethanol and glutaraldehyde (4:1 v:v) after being pretreated with 20% DMSO at 0[degree]C and slowly frozen to -20[degree]C, is also described. The choice of materials is described in detail. Electron micrographs of mouse tissue together with light micrographs of one micron sections stained to demonstrate enzyme activity are shown. The results compare favorably with other low-temperature and water-miscible methacrylate embedding methods. The techniques are discussed in the light of their usefulness as methods for the cytochemical investigation of plastic-embedded tissues. || ABSTRACT AUTHORS: Author