Raman-activated Single Cell Sorting and Analysis on Microfluidics Device

来源 :第八届全国微全分析系统学术会议、第三届全国微纳尺度生物分离分析学术会议暨第五届国际微化学与微系统学术会议 | 被引量 : 0次 | 上传用户:chen_gm
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  Raman-activated cell sorting (RACS) offers prospects to complement the widely applied fluorescence-activated 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 [1,2].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 [3].Attempts have been made to increase the cell sorting rate through the use of shorter Raman acquisition times on single cells[2].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 20um/s,and the sorting acuracy is about 70%.The sorting efficiency and accuracy improvement is still ongoing.In the future study,single cell genomics function will be integrated into the developed sorting system to realize complete procedure of single cell analysis.
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