Optimization of fluorophores for chemical tagging and immunohistochemistry of Drosophila neurons

优化用于果蝇神经元化学标记和免疫组织化学的荧光染料

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作者: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

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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