Biopolis Dresden Imaging Platform

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

Wong FK, Haffner C, Huttner WB, Taverna E

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.

Fig.3 taken from Wong et al, 2014.
  • Nat Protoc 2014 May 24;9(5):1170-82
  • 2014
  • Neurobiology
  • 24762784
  • PubMed

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