An update on molecular counting in fluorescence microscopy

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Quantitative assessment of protein complexes, such as receptor clusters in the context of cellular signalling, has become a pressing objective in cell biology. The advancements in the field of single molecule fluorescence microscopy have led to different approaches for counting protein copy numbers in various cellular structures. This has resulted in an increasing interest in robust calibration protocols addressing photophysical properties of fluorescent labels and the effect of labelling efficiencies. Here, we want to give an update on recent methods for protein counting with a focus on novel calibration protocols. In this context, we discuss different types of calibration samples and identify some of the challenges arising in molecular counting experiments. Some recently published applications offer potential approaches to tackle these challenges.

Bibliographic note

Funding Information: We would like to acknowledge helpful comments to the article from Klaus Yserentant. We are also grateful for funding from the Centre of Membrane Protein Research (COMPARE, Universities of Birmingham and Nottingham, UK) and the DFG (Deutsche Forschungsgemeinschaft , Germany, Grant HE4559/6-1 ). Publisher Copyright: © 2021 Elsevier Ltd


Original languageEnglish
Article number105978
Number of pages8
JournalThe International Journal of Biochemistry & Cell Biology
Early online date15 Apr 2021
Publication statusPublished - Jun 2021


  • Calibration standards, Oligomerization, Protein complexes, Quantitative microscopy

ASJC Scopus subject areas