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.
PLoS One. 2011 Mar 11;6(3):e17455. doi: 10.1371/journal.pone.0017455.
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.
从异质溶液中快速分离有活力的单细胞的技术为医学基因组学领域做出了重大贡献。目前仍然存在挑战,既要实现高效提取、分离和对异质溶液中单细胞的模式化处理,又要在这个过程中保持细胞的活力,因为对单细胞操作的控制程度有限。在这里,我们提出了一种基于微滴的方法,可以从异质细胞悬浮液(10%的目标细胞混合物)中分离和模式化单细胞,提取细胞的存活率达到 97.0±0.8%,并与非模式化对照相比,从分离的细胞中获得基因组信息。对细胞包封过程进行了实验和理论分析。使用分离的细胞,我们在 1000 个基因中鉴定出了 11 个干细胞标记物,并与对照进行了比较。与现有方法相比,这个自动化平台可实现高通量细胞操作,用于后续的基因组分析,操作步骤更少。