A Cryostat Method for the Extraction of Soluble Tissue Components

Abstract

ESTABLISHED techniques used to release the intracellular components of tissues either into solution or suspension include homogenization^1,2, mincing³, crushing⁴, and ultrasonic vibration⁵. The degree of difficulty in producing tissue homogenates varies for different tissues. In the case of cellular tissues containing little connective tissue almost complete cell disruption can be obtained with any of these techniques. Hogeboom et al.⁶ have shown that in unfiltered rat liver homogenates they were able to find only 7 × 10⁶ intact cells/g liver as compared with 268 × 10⁶ free nuclei. However, as the proportion of connective tissue increases, so does the work necessary to produce the same degree of cell disruption, and loss of labile cell contents generally follows as a result of prolonged exposure of the biological material to adverse physical conditions.