Open AccessOptimization of fluorophores for chemical tagging and immunohistochemistry of Drosophila neurons
Author(s) -
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
Publication year - 2018
Publication title -
plos one
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.99
H-Index - 332
ISSN - 1932-6203
DOI - 10.1371/journal.pone.0200759
Subject(s) - alexa fluor , fluorophore , drosophila melanogaster , cyanine , transgene , fluorescence , immunohistochemistry , chemistry , biology , microbiology and biotechnology , biophysics , biochemistry , gene , physics , quantum mechanics , immunology
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.