Microfluidic Device Based High Throughput Single Cell Sorting and Analysis

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  Raman-activated cell sorting (RACS) offers prospects to complement the widely applied fluorescenceactivated cell sorting.RACS enables the separation of cells according to their intrinsic chemical ‘fingerprint’ with minimal pre-treatment,thus cells are potentially viable after sorting.The isolated cells can then be further processed on a chip for attempted cultivation or DNA amplification.Spontaneous Raman based RACS has been demonstrated as a proof of concept in which SCRS combined with a microfluidic device and optical tweezers were applied to capture,analyse and sort suspended single cells in a low-throughput manner.Attempts have been made to increase the cell sorting rate through the use of shorter Raman acquisition times on single cells.We developed a RACS system based on microfluidics platform,which including Raman spectroscopy,532nm laser,cell switching system,sorting microfluidics chip and software.The Raman acquisition times of yeast and E.Coli cells were shorten to 10ms and 50ms separately,which is at least 100 times decreasing compared to traditional Raman system.The mechanism of RACS was shown in fig.1.The mixture of Cyanobacterium(Synechocystis sp.PCC6803) cells and fluorescent beads was sorted and counted to validate the sorting efficiency of the RACS system.2cells/s sorting effecieny was obtained when the flow rate is about 100um/s,and the sorting acuracy is about 70%.The sorting efficiency and accuracy improvement is still ongoing.Single cell genomics function will be integrated into the developed sorting system to realize overall procedure of single cell analysis.Microfluidic Device Based High Throughput Single Cell Sorting and Analysis
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