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  4. In vivo pulse labeling of isochronic cohorts of cells in the central nervous system using FlashTag
 
research article

In vivo pulse labeling of isochronic cohorts of cells in the central nervous system using FlashTag

Govindan, Subashika  
•
Oberst, Polina
•
Jabaudon, Denis
October 1, 2018
Nature Protocols

The tracing of neuronal cell lineages is critical to our understanding of cellular diversity in the CNS. This protocol describes a fluorescence birth-dating technique to label, track and isolate isochronic cohorts of newborn cells in the CNS in vivo in mouse embryos. Injection of carboxyfluorescein esters (CFSEs) into the cerebral ventricle allows pulse labeling of mitotic (M phase) ventricular zone (VZ) progenitors and their progeny across the CNS, a procedure we termed FlashTag. Specificity for M-phase apical progenitors is a result of the somata of these cells transiently contacting the ventricular wall during this cell-cycle phase, exposing them to CFSE injected into the cerebrospinal fluid. Using this approach, the developmental trajectory of progenitors and their daughter neurons can be tracked. Labeled cells can be imaged ex vivo or in fixed tissue, targeted for electrophysiological experiments or isolated using FACS for cell culture or (single-cell) RNA sequencing. Multiple embryos can be labeled within 30 min. The dye is retained for several weeks, allowing labeled cells to be identified postnatally. This protocol describes the labeling procedure using in utero injection, the isolation of live cells using FACS and the processing of labeled tissue for immunohistochemistry.

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Type
research article
DOI
10.1038/s41596-018-0038-1
Web of Science ID

WOS:000446087000008

Author(s)
Govindan, Subashika  
Oberst, Polina
Jabaudon, Denis
Date Issued

2018-10-01

Publisher

NATURE PUBLISHING GROUP

Published in
Nature Protocols
Volume

13

Issue

10

Start page

2297

End page

2311

Subjects

Biochemical Research Methods

•

Biochemistry & Molecular Biology

•

interkinetic nuclear migration

•

neural precursors

•

neurons

•

neocortex

•

proliferation

•

heterogeneity

•

epithelium

•

zones

Editorial or Peer reviewed

REVIEWED

Written at

EPFL

EPFL units
UPSUTER  
Available on Infoscience
December 13, 2018
Use this identifier to reference this record
https://infoscience.epfl.ch/handle/20.500.14299/152368
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