Quantifying Molecule Numbers in STED/RESOLFT Fluorescence Nanoscopy [chapter]

Jan Keller-Findeisen, Steffen J. Sahl, Stefan W. Hell
2020 Topics in applied physics  
Quantification of the numbers of molecules of interest in the specimen has emerged as a powerful capability of several fluorescence nanoscopy approaches. Carefully relating the measured signals from STED or RESOLFT scanning nanoscopy data to the contribution of a single molecule, reliable estimates of fluorescent molecule numbers can be obtained. To achieve this, higher-order signatures in the obtained photon statistics are analyzed, as arise from the antibunched nature of single-fluorophore
more » ... ngle-fluorophore emissions or in the signal variance among multiple on/off-switching cycles. In this chapter, we discuss the concepts and approaches demonstrated to date for counting molecules in STED/RESOLFT nanoscopy.
doi:10.1007/978-3-030-34413-9_7 fatcat:hpc3oiezzjdsvb2zeypu45xgva