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PLoS One. 2011 Mar 11;6(3):e17455. doi: 10.1371/journal.pone.0017455.

Drop-on-demand single cell isolation and total RNA analysis.

Author information

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.

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±0.8%), 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 identified 11 stem cell markers among 1000 genes and compare to the controls. This automated platform enabling high-throughput cell manipulation for subsequent genomic analysis employs fewer handling steps compared to existing methods.

PMID:
21412416
PMCID:
PMC3055874
DOI:
10.1371/journal.pone.0017455
[Indexed for MEDLINE]
Free PMC Article

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