Abstract: Using multilayer soft lithography technology, polydimethylsiloxane (PDMS) as the chip material, we made a novel microfluidic chip (60 mm×40 mm×4 mm) for single cell identification research. We enriched tumor stem cells in serumfree suspension culture of lung cancer cell A549 into single cell suspension.  RTPCR reaction liquid mixed into the chip, and cells were randomly distributed and reverse transcription PCR (RTPCR) in 2 048 micro chamber. The chip integrated in cell capture, separation, lysing and PCR reaction to achieve rapid identification of tumor stem cells. By counting the number of positive reaction, we could calculate the proportion of tumor stem cell in the total cells.

Key words: microfluidic chip, isolation and identification, one step, single cell, tumor stem cell