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 pr...

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Bibliographic Details
Published in:Journal of biotechnology 2001-04, Vol.86 (3), p.225-236
Main Authors: Dörre, Klaus, Stephan, Jens, Lapczyna, Markus, Stuke, Michael, Dunkel, Holger, Eigen, Manfred
Format: Article
Language:English
Subjects:
Biochemistry - instrumentation
Biochemistry - methods
Confocal set-up
Diffusion and flow
DNA sequencing
Elliptical volume
Fluorescence single molecule detection
Fluorescent Dyes - analysis
Image Processing, Computer-Assisted
Microstructure
Multi-element
Polymethyl Methacrylate
Spectrometry, Fluorescence - instrumentation
Spectrometry, Fluorescence - methods
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Summary: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.
ISSN:0168-1656
1873-4863
DOI:10.1016/S0168-1656(00)00415-6