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A waveguide imaging platform for live‐cell TIRF imaging of neurons over large fields of view
Author(s) -
Opstad Ida S.,
Ströhl Florian,
Fantham Marcus,
Hockings Colin,
Vanderpoorten Oliver,
Tartwijk Francesca W.,
Lin Julie Qiaojin,
Tinguely JeanClaude,
Dullo Firehun T.,
KaminskiSchierle Gabriele S.,
Ahluwalia Balpreet S.,
Kaminski Clemens F.
Publication year - 2020
Publication title -
journal of biophotonics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.877
H-Index - 66
eISSN - 1864-0648
pISSN - 1864-063X
DOI - 10.1002/jbio.201960222
Subject(s) - total internal reflection fluorescence microscope , live cell imaging , microscopy , hippocampal formation , neuroscience , microscope , fluorescence microscope , optics , biology , physics , cell , fluorescence , genetics
Abstract Large fields of view (FOVs) in total internal reflection fluorescence microscopy (TIRFM) via waveguides have been shown to be highly beneficial for single molecule localisation microscopy on fixed cells [1,2] and have also been demonstrated for short‐term live‐imaging of robust cell types [3‐5], but not yet for delicate primary neurons nor over extended periods of time. Here, we present a waveguide‐based TIRFM set‐up for live‐cell imaging of demanding samples. Using the developed microscope, referred to as the ChipScope , we demonstrate successful culturing and imaging of fibroblasts, primary rat hippocampal neurons and axons of Xenopus retinal ganglion cells (RGCs). The high contrast and gentle illumination mode provided by TIRFM coupled with the exceptionally large excitation areas and superior illumination homogeneity offered by photonic waveguides have potential for a wide application span in neuroscience applications.