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Reversible Chemical Reactions for Single‐Color Multiplexing Microscopy
Author(s) -
Brox Dominik,
Schwering Michael,
Engelhardt Johann,
Herten DirkPeter
Publication year - 2014
Publication title -
chemphyschem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.016
H-Index - 140
eISSN - 1439-7641
pISSN - 1439-4235
DOI - 10.1002/cphc.201402012
Subject(s) - multiplexing , microscopy , fluorescence , chemical imaging , fluorescence microscope , fluorescence lifetime imaging microscopy , spectral imaging , materials science , computer science , chemistry , nanotechnology , optics , physics , telecommunications , artificial intelligence , hyperspectral imaging
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.