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.
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June 2011
Research Article|
June 03 2011
Analysis of gene expression at the single-cell level using microdroplet-based microfluidic technology
Pascaline Mary;
Pascaline Mary
a)
1Experimental Soft and Condensed Matter Group,
SEAS
, Harvard, 58, Oxford Street, Cambridge, Massachusetts 02138, USA
2
MMN, UMR Gulliver, ESPCI
, 10 rue Vauquelin, 75005 Paris, France
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Luce Dauphinot;
Luce Dauphinot
3
ICM, INSERM, URMS975-CNRS UMR7225 Hôpital de la Pitié Salpétrière
, Bâtiment Pharmacie, 5ème étage 47 Bd. de l’Hôpital, 75013 Paris, France
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Nadège Bois;
Nadège Bois
3
ICM, INSERM, URMS975-CNRS UMR7225 Hôpital de la Pitié Salpétrière
, Bâtiment Pharmacie, 5ème étage 47 Bd. de l’Hôpital, 75013 Paris, France
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Marie-Claude Potier;
Marie-Claude Potier
3
ICM, INSERM, URMS975-CNRS UMR7225 Hôpital de la Pitié Salpétrière
, Bâtiment Pharmacie, 5ème étage 47 Bd. de l’Hôpital, 75013 Paris, France
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Vincent Studer;
Vincent Studer
b)
4Laboratoire de Neurobiologie, UMR 7637,
ESPCI
, 10 rue Vauquelin, 75005 Paris, France
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Patrick Tabeling
Patrick Tabeling
2
MMN, UMR Gulliver, ESPCI
, 10 rue Vauquelin, 75005 Paris, France
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Pascaline Mary
1,2,a)
Luce Dauphinot
3
Nadège Bois
3
Marie-Claude Potier
3
Vincent Studer
4,b)
Patrick Tabeling
2
1Experimental Soft and Condensed Matter Group,
SEAS
, Harvard, 58, Oxford Street, Cambridge, Massachusetts 02138, USA
2
MMN, UMR Gulliver, ESPCI
, 10 rue Vauquelin, 75005 Paris, France
3
ICM, INSERM, URMS975-CNRS UMR7225 Hôpital de la Pitié Salpétrière
, Bâtiment Pharmacie, 5ème étage 47 Bd. de l’Hôpital, 75013 Paris, France
4Laboratoire de Neurobiologie, UMR 7637,
ESPCI
, 10 rue Vauquelin, 75005 Paris, France
a)
Author to whom correspondence should be addressed. Tel.: (011)-1-617-496-0308. Electronic mail: [email protected].
b)
Present address: Laboratoire PCS, UMR CNRS 5091 Université Bordeaux 2, 146 rue Léo Saignat, 33077 Bordeaux, France.
Biomicrofluidics 5, 024109 (2011)
Article history
Received:
February 19 2011
Accepted:
April 28 2011
Citation
Pascaline Mary, Luce Dauphinot, Nadège Bois, Marie-Claude Potier, Vincent Studer, Patrick Tabeling; Analysis of gene expression at the single-cell level using microdroplet-based microfluidic technology. Biomicrofluidics 1 June 2011; 5 (2): 024109. https://doi.org/10.1063/1.3596394
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