Single cell detection in raw sample by a piece of membrane

Abstract

Micro/Nanofluidics has gain much attention recently, due to their potential on single cell detection and anal. Although much improvement has been made, the current chips still suffer from low throughput, limitation to microliter vol., laborious chip fabrication, and often require off-chip complex sample prepn. In this work, we report the single cell detection directly in raw samples by using a piece of com. membrane. Cell capture, concn., purifn., isolation, extn. and digital DNA amplification were successfully performed on one piece of membrane through simple syringe filtration and insitu digital LAMP (loop-mediated isothermal amplification). The membrane consisting of a high d. of micropores (25 um) on the top and nanochannels array (400 nm) on the bottom. The nanochannels were used for cell capture, concn. and purifn., while the micropore were for cell isolation. After filtration and in-situ digital LAMP, the micropore contg. target cell will generate bright fluorescence while micropores without target cell have very week fluorescence. By counting the no. of the bright pores, cell no. in the sample can be quantify precisely.

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