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Nat Protoc. 2014 May;9(5):1170-82. doi: 10.1038/nprot.2014.074. Epub 2014 Apr 24.

Microinjection of membrane-impermeable molecules into single neural stem cells in brain tissue.

Author information

1
Max Planck Institute of Molecular Cell Biology and Genetics (MPI-CBG), Dresden, Germany.

Abstract

This microinjection protocol allows the manipulation and tracking of neural stem and progenitor cells in tissue at single-cell resolution. We demonstrate how to apply microinjection to organotypic brain slices obtained from mice and ferrets; however, our technique is not limited to mouse and ferret embryos, but provides a means of introducing a wide variety of membrane-impermeable molecules (e.g., nucleic acids, proteins, hydrophilic compounds) into neural stem and progenitor cells of any developing mammalian brain. Microinjection experiments are conducted by using a phase-contrast microscope equipped with epifluorescence, a transjector and a micromanipulator. The procedure normally takes ∼2 h for an experienced researcher, and the entire protocol, including tissue processing, can be performed within 1 week. Thus, microinjection is a unique and versatile method for changing and tracking the fate of a cell in organotypic slice culture.

PMID:
24762784
DOI:
10.1038/nprot.2014.074
[Indexed for MEDLINE]

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