title: Ultrafast tissue staining with chemical tags.
abstract: Genetically encoded fluorescent proteins and immunostaining are widely used to detect cellular and subcellular structures in fixed biological samples. However, for thick or whole-mount tissue, each approach suffers from limitations, including limited spectral flexibility and lower signal or slow speed, poor penetration, and high background labeling, respectively. We have overcome these limitations by using transgenically expressed chemical tags for rapid, even, high-signal and low-background labeling of thick biological tissues. We first construct a platform of widely applicable transgenic Drosophila reporter lines, demonstrating that chemical labeling can accelerate staining of whole-mount fly brains by a factor of 100. Using viral vectors to deliver chemical tags into the mouse brain, we then demonstrate that this labeling strategy works well in mice. Thus this tag-based approach drastically improves the speed and specificity of labeling genetically marked cells in intact and/or thick biological samples.
authors.affiliation: Division of Neurobiology, Medical Research Council Laboratory of Molecular Biology, Cambridge CB2 0QH, United Kingdom.
authors.forename: Johannes
authors.initials: J
authors.lastname: Kohl
authors.affiliation: Division of Neurobiology, Medical Research Council Laboratory of Molecular Biology, Cambridge CB2 0QH, United Kingdom.
authors.forename: Julian
authors.initials: J
authors.lastname: Ng
authors.affiliation: Division of Neurobiology, Medical Research Council Laboratory of Molecular Biology, Cambridge CB2 0QH, United Kingdom.
authors.forename: Sebastian
authors.initials: S
authors.lastname: Cachero
authors.affiliation: Division of Neurobiology, Medical Research Council Laboratory of Molecular Biology, Cambridge CB2 0QH, United Kingdom.
authors.forename: Ernesto
authors.initials: E
authors.lastname: Ciabatti
authors.affiliation: Division of Neurobiology, Medical Research Council Laboratory of Molecular Biology, Cambridge CB2 0QH, United Kingdom.
authors.forename: Michael-John
authors.initials: MJ
authors.lastname: Dolan
authors.affiliation: Division of Neurobiology, Medical Research Council Laboratory of Molecular Biology, Cambridge CB2 0QH, United Kingdom.
authors.forename: Ben
authors.initials: B
authors.lastname: Sutcliffe
authors.affiliation: Division of Neurobiology, Medical Research Council Laboratory of Molecular Biology, Cambridge CB2 0QH, United Kingdom.
authors.forename: Adam
authors.initials: A
authors.lastname: Tozer
authors.affiliation: Division of Neurobiology, Medical Research Council Laboratory of Molecular Biology, Cambridge CB2 0QH, United Kingdom.
authors.forename: Sabine
authors.initials: S
authors.lastname: Ruehle
authors.affiliation: Division of Neurobiology, Medical Research Council Laboratory of Molecular Biology, Cambridge CB2 0QH, United Kingdom.
authors.forename: Daniel
authors.initials: D
authors.lastname: Krueger
authors.affiliation: Division of Neurobiology, Medical Research Council Laboratory of Molecular Biology, Cambridge CB2 0QH, United Kingdom.
authors.forename: Shahar
authors.initials: S
authors.lastname: Frechter
authors.affiliation: Division of Neurobiology, Medical Research Council Laboratory of Molecular Biology, Cambridge CB2 0QH, United Kingdom.
authors.forename: Tiago
authors.initials: T
authors.lastname: Branco
authors.affiliation: Division of Neurobiology, Medical Research Council Laboratory of Molecular Biology, Cambridge CB2 0QH, United Kingdom.
authors.forename: Marco
authors.initials: M
authors.lastname: Tripodi
authors.affiliation: Division of Neurobiology, Medical Research Council Laboratory of Molecular Biology, Cambridge CB2 0QH, United Kingdom jefferis@mrc-lmb.cam.ac.uk.
authors.forename: Gregory S X E
authors.initials: GS
authors.lastname: Jefferis
mesh_terms: D000818:Animals; D001921:Brain / Q000378:metabolism*; D004330:Drosophila; D005456:Fluorescent Dyes / Q000378:metabolism*; D008810:Mice, Inbred C57BL; D009474:Neurons / Q000166:cytology / Q000378:metabolism; D013194:Staining and Labeling / Q000379:methods*
chemical_list: D005456:Fluorescent Dyes
keywords: fluorescence microscopy; immunohistochemistry; neural circuits; protein labeling