TY - JOUR
T1 - Towards single molecule analysis in PDMS microdevices
T2 - From the detection of ultra low dye concentrations to single DNA molecule studies
AU - Ros, A.
AU - Hellmich, W.
AU - Duong, T.
AU - Anselmetti, D.
N1 - Funding Information:
Financial support from the Deutsche Forschungsgemeinschaft in the framework of the Sonderforschungsbereich 613 (Project number D2) as well as from the microchip UV LIF project (number An 370/1-1) are gratefully acknowledged. We thank Prof. Jürgen Brugger, EPFL (Switzerland), for technical support.
PY - 2004/8/26
Y1 - 2004/8/26
N2 - In this paper, we report on the performance of electrophoretical separation and laser-induced fluorescence (LIF) detection of dyes and fluorescently labeled biomolecules in poly(dimethylsiloxane) (PDMS) microdevices. The dyes fluorescein and fluorescein isothiocyanate (FITC) have been separated effectively in nM concentrations. Fluorescein injections gave linear concentration response in the range from 4 to 100 pM. As ultimate detection sensitivity, 100 fM injected fluorescein was obtained. Further, 100 fM injected fluorescein could be detected. This is to our knowledge the smallest electrokinetically injected dye concentration detected on a microchip. Injection studies of fluorescently labeled avidin revealed a theoretical detection limit of 25 nM for laser-induced fluorescence detection in good agreement with separations in glass chips. Furthermore, the injection of several and even one single DNA molecule using a PDMS cross injector has been demonstrated as well as free solution separation of λ- and T2-DNA (60 pM each) in periodically structured channels.
AB - In this paper, we report on the performance of electrophoretical separation and laser-induced fluorescence (LIF) detection of dyes and fluorescently labeled biomolecules in poly(dimethylsiloxane) (PDMS) microdevices. The dyes fluorescein and fluorescein isothiocyanate (FITC) have been separated effectively in nM concentrations. Fluorescein injections gave linear concentration response in the range from 4 to 100 pM. As ultimate detection sensitivity, 100 fM injected fluorescein was obtained. Further, 100 fM injected fluorescein could be detected. This is to our knowledge the smallest electrokinetically injected dye concentration detected on a microchip. Injection studies of fluorescently labeled avidin revealed a theoretical detection limit of 25 nM for laser-induced fluorescence detection in good agreement with separations in glass chips. Furthermore, the injection of several and even one single DNA molecule using a PDMS cross injector has been demonstrated as well as free solution separation of λ- and T2-DNA (60 pM each) in periodically structured channels.
KW - DNA migration
KW - Microchip separation
KW - Poly(dimethylsiloxane)
KW - Single molecule detection
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U2 - 10.1016/j.jbiotec.2004.04.020
DO - 10.1016/j.jbiotec.2004.04.020
M3 - Article
C2 - 15288941
AN - SCOPUS:3843068785
SN - 0168-1656
VL - 112
SP - 65
EP - 72
JO - Journal of Biotechnology
JF - Journal of Biotechnology
IS - 1-2
ER -