Fig. 8.

Analysis of the relation of TAMRA-positive cells to the clonotypic B cell. a Evaluation of the efficiency of cell sorting based on DNA internalization (Alu-TAMRA). b qPCR quantification of the B clone DNA (500 bp) content in sorted TAMRA+ and TAMRA− cells. The calibration curve was plotted using titration of purified B clone DNA (500 bp) and JH/FR3c primers. Dots denote amounts of the fragment in TAMRA+ and TAMRA– DNA. c Preparations of cells after internalization of the Flu or TAMRA Alu-DNA probe followed by FISH in a single preparation with the TAMRA-labeled 500 DNA fragment corresponding to the rearranged VDJ locus of the clonotypic B cell. 1, 2—Intact spheres with the Flu (1) and TAMRA (2) Alu-DNA probe. 3, 4—Sphere fragments. 5, 6—Cells of the fraction of free cells and destroyed sphere fragments. Arrows indicate the sites of specific hybridization of the probe’s DNA and cells captured the fluorochrome-labeled DNA probe. DAPI staining of nuclei