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Quantifying molecule numbers in STED/RESOLFT fluorescence nanoscopy

MPG-Autoren
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Hell,  Stefan W.
Optical Nanoscopy, Max Planck Institute for Medical Research, Max Planck Society;

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Zitation

Keller-Findeisen, J., Sahl, S. J., & Hell, S. W. (2020). Quantifying molecule numbers in STED/RESOLFT fluorescence nanoscopy. In Nanoscale Photonic Imaging (pp. 205-226). Berlin, Göttingen, Heidelberg: Springer, Cham.


Zitierlink: https://hdl.handle.net/21.11116/0000-0006-8B4D-8
Zusammenfassung
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 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.