Sample Preparation and Imaging Conditions Affect mEos3.2 Photophysics in Fission Yeast Cells.

ABSTRACT: Photoconvertible fluorescent proteins (PCFPs) are widely used in super-resolution microscopy and studies of cellular dynamics. However, our understanding of their photophysics is still limited, hampering their quantitative application. For example, we do not know the optimal sample preparation methods or imaging conditions to count protein molecules fused to PCFPs by single-molecule localization microscopy in live and fixed cells. We also do not know how the behavior of PCFPs in live cells compares with fixed cells. Therefore, we investigated how formaldehyde fixation influences the photophysical properties of the popular green-to-red PCFP mEos3.2 in fission yeast cells under a wide range of imaging conditions. We estimated photophysical parameters by fitting a three-state model of photoco

SUBMITTER: Sun M 

PROVIDER: S-EPMC7820738 | biostudies-literature | 2021 Jan

REPOSITORIES: biostudies-literature