Stereological method for estimating relative membrane surface area in freeze-fracture preparations of subcellular fractions.

Microsomal fractions from liver cells are a mixture of vesicles derived from a number of different cellular membranes. These can be differentiated on freeze-fracture preparations by their characteristic density of intramembranous particles. A stereological method is developed which allows the estimation of the relative membrane surface of the various membrane types by using freeze-fracture preparations. The sample is restricted to concave profiles without cast shadow. The numerical frequency distribution of vesicles with respect to particle density is determined. The estimation of relative surface area must consider the effects on sampling of (a) variable size distributions of vesicles and (b) loss of small profiles. Correction coefficients are derived which allow a differential vesicle count to be transformed into an estimate of relative membrane surface.