Long-term STED imaging of membrane packing and dynamics by exchangeable polarity-sensitive dyes

Understanding the plasma membrane nanoscale organization and dynamics in living cells requires microscopy techniques with high spatial and temporal resolution that permit for long acquisition times and allow for the quantification of membrane biophysical properties, such as lipid ordering. Among the...

Full description

Saved in:
Bibliographic Details
Published in:Biophysical reports 2021-12, Vol.1 (2), p.None-None, Article 100023
Main Authors: Carravilla, Pablo, Dasgupta, Anindita, Zhurgenbayeva, Gaukhar, Danylchuk, Dmytro I., Klymchenko, Andrey S., Sezgin, Erdinc, Eggeling, Christian
Format: Article
Language:English
Subjects:
Analytical chemistry
Biochemistry, Molecular Biology
Biophysics
Chemical Sciences
Life Sciences
Medicin och hälsovetenskap
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Understanding the plasma membrane nanoscale organization and dynamics in living cells requires microscopy techniques with high spatial and temporal resolution that permit for long acquisition times and allow for the quantification of membrane biophysical properties, such as lipid ordering. Among the most popular super-resolution techniques, stimulated emission depletion (STED) microscopy offers one of the highest temporal resolutions, ultimately defined by the scanning speed. However, monitoring live processes using STED microscopy is significantly limited by photobleaching, which recently has been circumvented by exchangeable membrane dyes that only temporarily reside in the membrane. Here, we show that NR4A, a polarity-sensitive exchangeable plasma membrane probe based on Nile red, permits the super-resolved quantification of membrane biophysical parameters in real time with high temporal and spatial resolution as well as long acquisition times. The potential of this polarity-sensitive exchangeable dye is showcased by live-cell real-time three-dimensional STED recordings of bleb formation and lipid exchange during membrane fusion as well as by STED-fluorescence correlation spectroscopy experiments for the simultaneous quantification of membrane dynamics and lipid packing that correlate in model and live-cell membranes.
ISSN:2667-0747
2667-0747
DOI:10.1016/j.bpr.2021.100023