Reversible chemical reactions for single-color multiplexing microscopy

Abstract Recent developments in biology demand an increasing number of simultaneously imaged structures with standard fluorescence microscopy. However, the number of multiplexed channels is limited for most multiplexing modalities, such as spectral multiplexing or fluorescence-lifetime imaging. We p...

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Bibliographic Details
Main Authors: Brox, Dominik (Author) , Schwering, Michael (Author) , Engelhardt, Johann (Author) , Herten, Dirk-Peter (Author)
Format: Article (Journal)
Language:English
Published: 17 April 2014
In: ChemPhysChem
Year: 2014, Volume: 15, Issue: 11, Pages: 2331-2336
ISSN:1439-7641
DOI:10.1002/cphc.201402012
Online Access:Verlag, lizenzpflichtig, Volltext: https://doi.org/10.1002/cphc.201402012
Verlag, lizenzpflichtig, Volltext: https://chemistry-europe.onlinelibrary.wiley.com/doi/abs/10.1002/cphc.201402012
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Author Notes:Dominik Brox, Michael Schwering, Johann Engelhardt, Dirk-Peter Herten
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Summary:Abstract Recent developments in biology demand an increasing number of simultaneously imaged structures with standard fluorescence microscopy. However, the number of multiplexed channels is limited for most multiplexing modalities, such as spectral multiplexing or fluorescence-lifetime imaging. We propose extending the number of imaging channels by using chemical reactions, controlling the emissive state of fluorescent dyes. As proof of concept, we reversibly switch a fluorescent copper sensor to enable successive imaging of two different structures in the same spectral channel. We also show that this chemical multiplexing is orthogonal to existing methods. By using two different dyes, we combine chemical with spectral multiplexing for the simultaneous imaging of four different structures with only two spectrally different channels. We characterize and discuss the approach and provide perspectives for extending imaging modalities in stimulated emission depletion microscopy, for which spectral multiplexing is technically demanding.
Item Description:Gesehen am 27.08.2020
Physical Description:Online Resource
ISSN:1439-7641
DOI:10.1002/cphc.201402012