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Optimization of fluorophores for chemical tagging and immunohistochemistry of Drosophila neurons.

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.

SUBMITTER: Meissner GW

PROVIDER: S-EPMC6093644 | biostudies-literature | 2018

REPOSITORIES: biostudies-literature

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Optimization of fluorophores for chemical tagging and immunohistochemistry of Drosophila neurons.

Meissner Geoffrey W GW Grimm Jonathan B JB Johnston Rebecca M RM Sutcliffe Ben B Ng Julian J Jefferis Gregory S X E GSXE Cachero Sebastian S Lavis Luke D LD Malkesman Oz O

PloS one 20180815 8

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 ...[more]

PMID: 30110347

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