| Description |
This dissertation studies detection-based methods to increase the estimation precision of single point-source emitters in the field of 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. These theoretical calculation and simulations are demonstrated through the use of two proof-of-concept experiments utilizing a modified Mach-Zehnder interferometer. The first methodology improves the localization precision of a single nanoparticle over the theoretical limit for an Airy-disk point-spread function by using self-interference to spatially modulate the recorded point-spread function. Experimental analysis demonstrates an improvement factor of ~3 to 5 over conventional localization methods. A related method employs the phase induced onto the Fourier domain signal due to path length differences in the Mach-Zehnder interferometer to improve localization precision. The localization capability of a modified Fourier domain signal generated by self-interference is utilized to yield a two-fold improvement in the localization precision for a given number of photons compared to a standard Gaussian intensity distribution of the corresponding point-spread function. |
