Format

Send to

Choose Destination
Proc Natl Acad Sci U S A. 2014 Sep 9;111(36):E3805-14. doi: 10.1073/pnas.1411087111. Epub 2014 Aug 25.

Ultrafast tissue staining with chemical tags.

Author information

1
Division of Neurobiology, Medical Research Council Laboratory of Molecular Biology, Cambridge CB2 0QH, United Kingdom.
2
Division of Neurobiology, Medical Research Council Laboratory of Molecular Biology, Cambridge CB2 0QH, United Kingdom jefferis@mrc-lmb.cam.ac.uk.

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.

KEYWORDS:

fluorescence microscopy; immunohistochemistry; neural circuits; protein labeling

PMID:
25157152
PMCID:
PMC4246963
DOI:
10.1073/pnas.1411087111
[Indexed for MEDLINE]
Free PMC Article

Supplemental Content

Full text links

Icon for HighWire Icon for PubMed Central
Loading ...
Support Center