Analysis of gene expression at the single-cell level using microdroplet-based microfluidic technology
Biomicrofluidics. 2011-06-01; 5(2): 024109
DOI: 10.1063/1.3596394
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1. Biomicrofluidics. 2011 Jun;5(2):24109. doi: 10.1063/1.3596394. Epub 2011 Jun 3.
Analysis of gene expression at the single-cell level using microdroplet-based
microfluidic technology.
Mary P, Dauphinot L, Bois N, Potier MC, Studer V, Tabeling P.
In the present work, we have measured the messenger RNA expression of specific
genes both from total RNA and cells encapsulated in droplets. The microfluidic
chip introduced includes the following functionalities: RNA∕cell encapsulation,
lysis, reverse transcription and real-time polymerase chain reaction. We have
shown that simplex and duplex gene expression measurements can be carried out
over a population of 100 purified RNA samples encapsulated simultaneously in 2 nl
droplets in less than 2 h. An analysis of 100 samples containing one to three
cells has shown excellent consistency with standard techniques regarding average
values. The cell-to-cell distributions of the E-cadherin expression suggest
fluctuations on the order of 80% in the number of transcripts, which is highly
consistent with the general findings from the literature. A mathematical model
has also been introduced to strengthen the interpretation of our results. The
present work paves the way for the systematic acquisition of such information in
biological and biomedical studies.
DOI: 10.1063/1.3596394
PMCID: PMC3124518
PMID: 21716808