Multiple-Emitter Super-resolution Imaging using the Alternating Descent Conditional Gradient Method

Sammanfattning: This thesis examines the state-of-the-art 2D super-resolution technique alternating descent conditional gradient (ADCG) method's ability to accurately localize fluorophores in diffraction-limited single molecule images (SMI) and analyze the impact of pre-processing and post-processing modules on ADCG's fluorophore localization. A synthetic dataset obtained from the 2013 Grand Challenge localization microscopy and a temporally linked dataset obtained from an unpublished set of Optical DNA mapping experiments performed by Jonathan Jeffet at the NanoBioPhotonix Lab at Tel-Aviv University were initially segmented to extract their noise parameters. Subsequently, the fluorophores in both datasets were localized on the order of nanometers and tens of nanometers, respectively, using ADCG. Additionally, leveraging intrinsic fluorophore behavior, as detailed in Jaqaman et al., improved the specificity of the temporally linked localizations. Moreover, by time averaging the fluorophore positions, as detailed in Jeffet et al., the precision of the temporally linked localizations was improved by a factor of 2. Consequently, it is concluded that ADCG can rapidly generate super-resolution images, robustly track single molecules, and generate DNA barcodes for diffraction-limited SMIs in an automatable fashion.