Sarkar, Susanta K.Sullivan, Neal P.Cannataro, Frank2007-01-032022-02-032007-01-032022-02-032015https://hdl.handle.net/11124/171422015 Spring.Includes illustrations (some color).Includes bibliographical references (pages 40-45).We mostly think and communicate science at the single-molecule level, but do experiments at the ensemble level. This gap between our thinking and experiments has been closing with the advent of single-molecule techniques nearly twenty years ago. Single-molecule techniques have proved very useful in probing individual molecules and heterogeneities, and have provided unprecedented insights into previously inaccessible scientific knowledge. Studying molecules one by one using single-molecule techniques has become very useful in revealing insights into mechanistic details hidden in ensemble experiments. I have built a total internal reflection fluorescence (TIRF) microscope with single-molecule sensitivity to study fluorescent molecules. The microscope is modular and multifunctional with capabilities including a wide excitation wavelength range (400-2000 nm) both in objective-type and prism-type TIRF configurations, localization accuracy ~1 nm, temporal resolution ~1 ms, and alternate laser excitation. As an application, we have measured high-throughput photoluminescent spectra of individual CdSe quantum dots.born digitalmasters thesesengCopyright of the original work is retained by the author.total internal reflection fluorescencefluorescencesingle moleculeoptomechanicalmicroscopyspectroscopyTotal internal reflection (Optics)MoleculesFluorescenceMicroscopySpectrum analysisDevelopment of single molecule total internal reflection fluorescence microscopeText