https://hal.archives-ouvertes.fr/hal-01604987/document

Size-dependent DNA Mobility in Cytoplasm and Nucleus

The photostability of fluorescent dyes is of crucial importance for the statistical accuracy of single-molecule detection (SMD) and for the image quality of scanning confocal microscopy. Concurrent results for the photostability were obtained by two different experimental techniques. First, the photostabilities of several coumarin and rhodamine derivatives in aqueous solution were obtained by monitoring the steady-state fluorescence decay in a quartz cell. Furthermore, an epi-illuminated microscope, continuous wave (CW) excitation at 514.5 nm, and fluorescence correlation spectroscopy (FCS) with a newly developed theory were used to study the photobleaching characteristics of rhodamines under conditions used for SMD. Depending on the rhodamine structure, the probability of photobleaching, pb, is in the order of 10-6−10-7 for irradiances below 103 W/cm2. However, a considerable increase of pb for irradiances above this level was observed which can only be described by photobleaching reactions from higher e...

Photobleaching of Fluorescent Dyes under Conditions Used for Single-Molecule Detection: Evidence of Two-Step Photolysis.

Ever since their introduction two decades ago, single-molecule (SM) fluorescence methods have matured and branched out to address numerous biological questions, which were inaccessible via ensemble measurements. Among the current arsenal, SM fluorescence techniques have capabilities of probing the dynamic interactions of nucleic acids and proteins via Forster (fluorescence) resonance energy transfer (FRET), tracking single particles over microns of distances, and deciphering the rotational motion of multisubunit systems. In this exciting era of transitioning from in vitro to in vivo and in situ conditions, it is anticipated that SM fluorescence methodology will become a common tool of molecular biology.

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Advances in Single-Molecule Fluorescence Methods for Molecular Biology

Photobleaching recovery and anisotropy decay of green fluorescent protein GFP-S65T in solution and cells: cytoplasmic viscosity probed by green fluorescent protein translational and rotational diffusion.

https://www.cell.com/trends/biochemical-sciences/pdf/S0968-0004(01)02003-5.pdf

Solute and macromolecule diffusion in cellular aqueous compartments

https://pure.uva.nl/ws/files/969079/1628_17038y.pdf

C.A.G.O. Varma

Papers

Influence of the triplet excited state on the photobleaching kinetics of fluorescein in microscopy