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URL: https://dx.doi.org/DOI:10.17504/protocols.io.qf6dtre

Authors: Geoffrey W. Meissner, Jonathan B. Grimm, Rebecca M. Johnston, Ben Sutcliffe, Julian Ng, Gregory S.X.E. Jefferis, Sebastian Cachero, Luke D. Lavis, Oz Malkesman

Summary: 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.

Affiliations: Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, Virginia, USA, Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, Virginia, USA, Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, Virginia, USA, Division of Neurobiology, Medical Research Council Laboratory of Molecular Biology, Cambridge CB2 0QH, United Kingdom, Division of Neurobiology, Medical Research Council Laboratory of Molecular Biology, Cambridge CB2 0QH, United Kingdom, Division of Neurobiology, Medical Research Council Laboratory of Molecular Biology, Cambridge CB2 0QH, United Kingdom, Division of Neurobiology, Medical Research Council Laboratory of Molecular Biology, Cambridge CB2 0QH, United Kingdom, Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, Virginia, USA, Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, Virginia, USA

External URL: https://www.janelia.org/project-team/flylight/protocols

Version: 1

Publication Date: 2018

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