Carl Ebeling Thesis Defense 12/05/14

Thesis Defense

Carl Ebeling

Friday, December 5, 2014
2:00PM (210 ASB)

Title: Increasing Estimation Precision in Localization Microscopy

Localization microscopy is a novel method allowing for the localization of optical point-source emitters below the Abbe diffraction limit of optical microscopy. This is accomplished by optically controlling the active, or bright, state of individual molecules within a sample. The use of time multiplexing of the active state allows for the temporal and spatial isolation of single point-source emitters. Isolating individual sources within a sample allows for statistical analysis on their emission point spread function profile, and the spatial coordinates of the point-source may be discerned below the optical response of the microscope system. Localization microscopy enables the identification of individual point-source emitter locations approximately an order of magnitude below standard, diffraction-limited optical techniques.

The precision of localization microscopy methods is limited by the statistical uncertainty in which the location of these sources may be estimated. By utilizing a detection-based interferometer, an interference pattern may be super-imposed over the emission signal. Theoretical analysis and Monte-Carlo simulations by means of Fisher information theory demonstrate that the incorporation of a modulation structure over the emission signal allow for a more precise estimation when compared to conventional localization methods for the same number of recorded photons. Experimental demonstration and verification of these ideas are presented and will be discussed for two separate imaging modalities.

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