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Study on Separation of Sperm and Epithelial Cells in a Microfluidic Chip
Ou Yuan1,2#, Liu Weiran3#, Dong Junlei4, Li Caixia1, Jin Xianting5, Liu Lin4, Zhao Xingchun1, Ye Jian1*
1Institute of Forensic Science, Ministry of Public Security, Beijing 100038, China; 2School of Life Science,Beijing Institute of Technology, Beijing 100081, China; 3Department of Biomedical Engineering,Tsinghua University,
Abstract: To develop the method for separation of sperm and epithelial cells in a microfluidic chip， the glass- PDMS chip， which is easy to manufacture， was selected to separation of sperm and epithelial cells. Buffer was added to the inlet (7 μL) and outlet reservoirs (10 μL) prior to sample addition， then a sperm/epithelial cell mixture (2 μL) was added to the inlet reservoir. After a minimum of 8 min of “settling time”， gravity-induced flow was initiated by removing 3 μL of buffer from the outlet reservoir， then the inlet reservoirs was supplemented with 1μL buffer every 5min. Following the desired separation time， product was removed from the outlet reservoir via pipet. Nuclease was used to remove free DNA， followed by extraction， amplification and electrophoretic separation/detection. By using microfabricated chip based gravity-induced flow mechanism， sperm can be separated from mixture in 30 min. No epithelial cells were observed passing through the channel during separation. After removing free DNA by using nuclease， a single and complete sperm STR profile can be obtained. Compared with the conventional differential extraction， this method can largely save test time and has a certain value for the analysis of forensic evidence in sexual assault cases.
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