Highly efficient single molecule detection in microstructures.
For DNA single molecule sequencing, the complete detection of all dye-labeled monomers which are cleaved off during the sequencing reaction is an essential requirement. In this work we address the feasibility of single molecule detection in microstructures with a confocal multi element set-up. We present statistical data on single molecule recognition and explain a refined data evaluation technique for single molecule burst analysis. From these data the signal-to-noise ratio in microstructures is evaluated as well as the overall detection efficiency. So far, detection efficiencies of single molecule events of up to 60% have been shown in microstructures.
Max-Planck-Institut für biophysikalische Chemie, Abteilung Biochemische Kinetik, Am Fassberg 11, D-37077 Göttingen, Germany. kdoerre@gwdg.de
K
K
Dörre
J
J
Stephan
M
M
Lapczyna
M
M
Stuke
H
H
Dunkel
M
M
Eigen
D001671:Biochemistry / Q000295:instrumentation* / Q000379:methods*; D005456:Fluorescent Dyes / Q000032:analysis*; D007091:Image Processing, Computer-Assisted; D019904:Polymethyl Methacrylate; D013050:Spectrometry, Fluorescence / Q000295:instrumentation / Q000379:methods
D005456:Fluorescent Dyes; D019904:Polymethyl Methacrylate