A microsample collection device for electrostatically sorted cells or particles and its preparative use for biochemical analysis

Abstract

Biochemical analysis of proteins extracted from mammalian cell nuclei which were sorted according to DNA content was greatly simplified using a special collection device to recover sorted nuclei. The device was developed for use with a sorting flow cytometer to provide sufficient numbers of particles to achieve adequate sensitivity and resolution while minimizing sample degradation and loss. The design, evaluation and application of the collection device were described with emphasis on sample preparation. Nuclear populations G0 + G1, S, and G2 + M, were sorted form regenerating rat liver tissue preparations and collected separately in the special chambers. Nuclear proteins were extracted and analyzed by polyacrylamide gel isoelectric focusing or capillary isotachophoresis.