The freeze substitution histological technique allows the indirect light microscopicobservation and the quantitative evaluation of ice crystal size in frozen tissues. Theuseofchemical fixatives in substitu ting fluids improves morphological and histochemical preservation of the tissue. Fixation conditions become important since this step can introduce modifications in crystal sizes . Effects of temperature on: a) diffusion rate of fixing solution in tissue, b) recrystallization rate of ice in frozen beef and c) variations of the froze~ water fraction were ana lyzed, establishing that 1sothermal freeze fixation constitutes an appropriate method for histological observation. Fixing at temperatures lower than that of the sample, in an attempt to reduce recrystallization effects , involves an increase of the frozen water fraction i.n the tissue leading to modifications in size of ice crystals and to changes in the histological structure.
Martino, M. N. and Zaritzky, N. E.
"Fixing Conditions in the Freeze Substitution Technique for Light Microscopy Observation of Frozen Beef Tissue,"
1, Article 3.
Available at: http://digitalcommons.usu.edu/foodmicrostructure/vol5/iss1/3