A waveguide imaging platform for live-cell TIRF imaging of neurons over large fields of view

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 and have also been demonstrated for short-term live-imaging of robust cell types, but not yet for...

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Published inbioRxiv
Main Authors Opstad, Ida S, Strohl, Florian, Fantham, Marcus, Hockings, Colin, Vanderpoorten, Oliver, Francesca Van Tartwijk, Lin, Julie Qiaojin, Tinguely, Jean-Claude, Dullo, Firehun T, Kaminski-Schierle, Gabriele S, Balpreet Singh Ahluwalia, Kaminski, Clemens F
Format Paper
LanguageEnglish
Published Cold Spring Harbor Cold Spring Harbor Laboratory Press 13.12.2019
Subjects
Axons
Fibroblasts
Fluorescence microscopy
Hippocampus
Illumination
Microscopy
Nervous system
Neurons
Retina
Retinal ganglion cells
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Summary: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 and have also been demonstrated for short-term live-imaging of robust cell types, 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 (RGC). 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.
DOI:10.1101/2019.12.13.874545