Optimization of fluorophores for chemical tagging and immunohistochemistry of Drosophila neurons.
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Peer-reviewed
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Abstract
The use of genetically encoded 'self-labeling tags' with chemical fluorophore ligands enables rapid labeling of specific cells in neural tissue. To improve the chemical tagging of neurons, we synthesized and evaluated new fluorophore ligands based on Cy, Janelia Fluor, Alexa Fluor, and ATTO dyes and tested these with recently improved Drosophila melanogaster transgenes. We found that tissue clearing and mounting in DPX substantially improves signal quality when combined with specific non-cyanine fluorophores. We compared and combined this labeling technique with standard immunohistochemistry in the Drosophila brain.
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Keywords
Animals, Animals, Genetically Modified, Brain, Drosophila melanogaster, Female, Fluorescent Dyes, Immunohistochemistry, Microscopy, Confocal, Molecular Structure, Neurons, Staining and Labeling
Journal Title
PLoS One
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1932-6203
1932-6203
1932-6203
Volume Title
13
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PLOS
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European Research Council (649111)
This work was supported by Howard Hughes Medical Institute (https://www.hhmi.org), the Medical Research Council (https://mrc.ukri.org; MRC file reference U105188491) and a European Research Council (https://erc.europa.eu) Consolidator grant (649111) to G.S.X.E.J., and a Royal Society (https://royalsociety.org) Dorothy Hodgkin Fellowship to S.C.