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Advanced spinning disk‐TIRF microscopy for faster imaging of the cell interior and the plasma membrane
Author(s) -
ZOBIAK BERND,
FAILLA ANTONIO VIRGILIO
Publication year - 2018
Publication title -
journal of microscopy
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.569
H-Index - 111
eISSN - 1365-2818
pISSN - 0022-2720
DOI - 10.1111/jmi.12626
Subject(s) - total internal reflection fluorescence microscope , microscopy , membrane , plasma , microscope , fluorescence microscope , spinning , resolution (logic) , live cell imaging , reflection (computer programming) , materials science , biophysics , chemistry , optics , fluorescence , cell , physics , biology , computer science , biochemistry , artificial intelligence , quantum mechanics , composite material , programming language
Summary Understanding the cellular processes that occur between the cytosol and the plasma membrane is an important task for biological research. Till now, however, it was not possible to combine fast and high‐resolution imaging of both the isolated plasma membrane and the surrounding intracellular volume. Here, we demonstrate the combination of fast high‐resolution spinning disk (SD) and total internal reflection fluorescence (TIRF) microscopy for specific imaging of the plasma membrane. A customised SD‐TIRF microscope was used with specific design of the light paths that allowed, for the first time, live SD‐TIRF experiments at high acquisition rates. A series of experiments is shown to demonstrate the feasibility and performance of our setup.