drop-on-demand single cell isolation and total rna analysisdrop-on-demand单细胞隔离和总rna分析.pdfVIP

Drop-on-Demand Single Cell Isolation and Total RNA Analysis 1 1 2 2 3 Sangjun Moon , Yun-Gon Kim , Lingsheng Dong , Michael Lombardi , Edward Haeggstrom , Roderick V. 4 2 1,2,5 Jensen , Li-Li Hsiao *, Utkan Demirci * 1 Demirci Bio-Acoustic-MEMS in Medicine (BAMM) Laboratory, Center for Bioengineering, Brigham and Women’s Hospital, Harvard Medical School, Boston, Massachusetts, United States of America, 2 Renal Division, Brigham and Women’s Hospital, Harvard Medical School, Boston, Massachusetts, United States of America, 3 Electronics Research Laboratory, Department of Physics, University of Helsinki, Helsinki, Finland, 4 Department of Biological Sciences, Virginia Tech, Blacksburg, Virginia, United States of America, 5 Harvard-MIT Division of Health Sciences and Technology, Massachusetts Institute of Technology, Cambridge, Massachusetts, United States of America Abstract Technologies that rapidly isolate viable single cells from heterogeneous solutions have significantly contributed to the field of medical genomics. Challenges remain both to enable efficient extraction, isolation and patterning of single cells from heterogeneous solutions as well as to keep them alive during the process due to a limited degree of control over single cell manipulation. Here, we present a microdroplet based method to isolate and pattern single cells from heterogeneous cell suspensions (10% target cell mixture), preserve viability of the extracted cells (97.0 60.8%), and obtain genomic information from isolated cells compared to the non-patterned controls. The cell encapsulation process is both experimentally and theoretically analyzed. Using the isolated cells, we ide