Damage-less sorting system for live cells

Abstract

Rapid development of modern medicine requires the system which can acquire living cells, especially on stem cells and human embryonic stem (hES) cells in good condition. Taking advantage of its proprietary technologies in fiber optics and mechatronics, Furukawa Electric has developed and commoditized a compacted flowcytometer (PERFLOW^TM) that features damage-less sorting for the first time in the world. Unlike the conventional optical lens system configuration, PERFLOW eliminates air layer intervention allowing high-sensitivity measurement through directly connecting optical fibers for both laser light and fluorescent and scattered light receptions to the flow cell. This optics structure enables us to obtain new information from transmission light instead of forward scatter light conventionally used. Measurement results of human peripheral blood mononuclear cells as well as lymphocyte subsets using our transmission / side scatter light system showed a distribution pattern similar to that obtained with conventional forward scatter / side scatter system. Furthermore, measurement of HeLa cells synchronized in each cell-cycle phase with transmission light signals displayed an increase in size consistent with their actual size (G2/M>S>G1), indicating that transmission light signals have a higher accuracy than forward scatter signals. Our PERFLOW system has a performance of damage-less sorting of single living cell. Megakaryocyte, one of the most delicate blood cells which has never been sorted alive by conventional sorter so far, was alive three days after sorting with our PERFLOW system. Additionally, single neural crest stem cell was also successfully sorted using the PERFLOW system and formed a colony after 20 days culture in vitro. Taken together, these results demonstrate that PERFLOW is a compacted flowcytometer with a high performance in cell analysis and sorting. Through further improvement, it will provide more novel information and become a useful tool of medical researches.