Single cell gene expression profiling of cell lines and murine peripheral blood mononuclear cells (PBMCs)

To demonstrate the efficacy of our developed Automated Live-imaging and cell Picking System for downstream RNA sequencing (ALPS&RNA-seq), we isolated single cells of three cell lines (murine T cell, murine leukemia cell, and murine hematopoietic progenitor cell) and of murine peripheral blood mononuclear cells (PBMCs). The detemined profiles of the three cell lines were different, and the profiles of the PBMSc showed three subpopulations which were determined as CD4+ T cells, CD8+ T cells, and B cells based on the marker genes. Overall design: Four 96-well plates containing single cells were isolated from a cell line mixture of murine leukemia cells, T cells, and hematopoietic progenitor cells by ALPS with random selection mode (3mix-random). Four plates of 36 leukemia cells, 36 T cells, 24 hematopoietic progenitor cells in order were isolated by ALPS with target selection mode (3mix-target). Two plates of 36 leukemia cells, 36 T cells, 24 hematopoietic progenitor cells in order were sorted by FACS (3mix-FACS). Sixteen plates of leukemia cells, T cells, and hematopoietic progenitor cells were isolated by ALPS after time-lapse imaging (3mix-timelapse). Eight plates of PBMCs of C57BL/6J mice were isolated by ALPS after time-lapse imaging (PBMC-timelapse).