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Real-time Monitoring and Manipulation of Single Bio-Molecules in Free Solution.

DE2006850082

Publication Date	2005
Personal Author	Li, H. W.
Page Count	132
Abstract	The observation and manipulation of single biomolecules allow their dynamic behaviors to be studied to provide insight into molecular genetics, biochip assembly, biosensor design, DNA biophysics. In a PDMS/glass microchannel, a nonuniform electroosmotic flow (EOF) was created. By using a scanning confocal fluorescence microscope and total internal-refletion fluorescence microscope (TIRFM), we demonstrated that negatively charged DNA molecules were focused by the nonuniform EOF into a thin layer at the glass surface. This phenomenon was applied to selectively detect target DNA molecules without requiring the separation of excessive probes and can be applied continuously to achieve high throughput. A variable-angle-TIRFM was constructed for imaging single DNA molecule dynamics at a solid/liquid interface. Implications we have are that the measured intensities cannot be used directly to determine the distances of molecules from the surface and the experimental counting results depict the distance-dependent dynamics of molecules near the surface.
Keywords	Biomolecules Molecular biology Adenines Biophysics Chromatography Conformational changes Deoxyribonucleic acid Electrostatics Excitation Fluorescence Genetics Isoalloxazines Monitoring Pyrimidines
Source Agency	Technical Information Center Oak Ridge Tennessee
Corporate Authors	Iowa State Univ., Ames.; Department of Energy, Washington, DC.
Supplemental Notes	Sponsored by Department of Energy, Washington, DC.
Document Type	Technical Report
Title Note	Doctoral Thesis.
NTIS Issue Number	200615

Real-time Monitoring and Manipulation of Single Bio-Molecules in Free Solution.

Real-time Monitoring and Manipulation of Single Bio-Molecules in Free Solution.
DE2006850082

Publication Date	2005
Personal Author	Li, H. W.
Page Count	132
Abstract	The observation and manipulation of single biomolecules allow their dynamic behaviors to be studied to provide insight into molecular genetics, biochip assembly, biosensor design, DNA biophysics. In a PDMS/glass microchannel, a nonuniform electroosmotic flow (EOF) was created. By using a scanning confocal fluorescence microscope and total internal-refletion fluorescence microscope (TIRFM), we demonstrated that negatively charged DNA molecules were focused by the nonuniform EOF into a thin layer at the glass surface. This phenomenon was applied to selectively detect target DNA molecules without requiring the separation of excessive probes and can be applied continuously to achieve high throughput. A variable-angle-TIRFM was constructed for imaging single DNA molecule dynamics at a solid/liquid interface. Implications we have are that the measured intensities cannot be used directly to determine the distances of molecules from the surface and the experimental counting results depict the distance-dependent dynamics of molecules near the surface.
Keywords	Biomolecules Molecular biology Adenines Biophysics Chromatography Conformational changes Deoxyribonucleic acid Electrostatics Excitation Fluorescence Genetics Isoalloxazines Monitoring Pyrimidines
Source Agency	Technical Information Center Oak Ridge Tennessee
Corporate Authors	Iowa State Univ., Ames.; Department of Energy, Washington, DC.
Supplemental Notes	Sponsored by Department of Energy, Washington, DC.
Document Type	Technical Report
Title Note	Doctoral Thesis.
NTIS Issue Number	200615