Microinjection of membrane-impermeable molecules into single neural stem cells in brain tissue.
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.
Max Planck Institute of Molecular Cell Biology and Genetics (MPI-CBG), Dresden, Germany.
Fong Kuan
FK
Wong
Max Planck Institute of Molecular Cell Biology and Genetics (MPI-CBG), Dresden, Germany.
Christiane
C
Haffner
Max Planck Institute of Molecular Cell Biology and Genetics (MPI-CBG), Dresden, Germany.
Wieland B
WB
Huttner
Max Planck Institute of Molecular Cell Biology and Genetics (MPI-CBG), Dresden, Germany.
Elena
E
Taverna
D000818:Animals; D001921:Brain / Q000166:cytology*; D005289:Ferrets; D005453:Fluorescence; D051379:Mice; D008845:Microinjections / Q000379:methods*; D008846:Micromanipulation / Q000379:methods; D008858:Microscopy, Phase-Contrast; D058953:Neural Stem Cells / Q000378:metabolism*