TY - JOUR
T1 - Modeling Non-additive Effects in Neighboring Chemically Identical Fluorophores
AU - Saurabh, Ayush
AU - Niekamp, Stefan
AU - Sgouralis, Ioannis
AU - Pressé, Steve
N1 - Publisher Copyright:
© 2022 American Chemical Society.
PY - 2022/6/16
Y1 - 2022/6/16
N2 - Quantitative fluorescence analysis is often used to derive chemical properties, including stoichiometries, of biomolecular complexes. One fundamental underlying assumption in the analysis of fluorescence data whether it be the determination of protein complex stoichiometry by super-resolution, or step-counting by photobleaching, or the determination of RNA counts in diffraction-limited spots in RNA fluorescence in situ hybridization (RNA-FISH) experimentsis that fluorophores behave identically and do not interact. However, recent experiments on fluorophore-labeled DNA origami structures such as fluorocubes have shed light on the nature of the interactions between identical fluorophores as these are brought closer together, thereby raising questions on the validity of the modeling assumption that fluorophores do not interact. Here, we analyze photon arrival data under pulsed illumination from fluorocubes where distances between dyes range from 2 to 10 nm. We discuss the implications of non-additivity of brightness on quantitative fluorescence analysis.
AB - Quantitative fluorescence analysis is often used to derive chemical properties, including stoichiometries, of biomolecular complexes. One fundamental underlying assumption in the analysis of fluorescence data whether it be the determination of protein complex stoichiometry by super-resolution, or step-counting by photobleaching, or the determination of RNA counts in diffraction-limited spots in RNA fluorescence in situ hybridization (RNA-FISH) experimentsis that fluorophores behave identically and do not interact. However, recent experiments on fluorophore-labeled DNA origami structures such as fluorocubes have shed light on the nature of the interactions between identical fluorophores as these are brought closer together, thereby raising questions on the validity of the modeling assumption that fluorophores do not interact. Here, we analyze photon arrival data under pulsed illumination from fluorocubes where distances between dyes range from 2 to 10 nm. We discuss the implications of non-additivity of brightness on quantitative fluorescence analysis.
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U2 - 10.1021/acs.jpcb.2c01889
DO - 10.1021/acs.jpcb.2c01889
M3 - Article
AN - SCOPUS:85131966269
SN - 1520-6106
VL - 126
SP - 4216
EP - 4225
JO - Journal of Physical Chemistry B
JF - Journal of Physical Chemistry B
IS - 23
ER -