Live imaging of Drosophila melanogaster embryonic hemocyte migrations.
Many studies address cell migration using in vitro methods, whereas the physiologically relevant environment is that of the organism itself. Here we present a protocol for the mounting of Drosophila melanogaster embryos and subsequent live imaging of fluorescently labeled hemocytes, the embryonic macrophages of this organism. Using the Gal4-uas system we drive the expression of a variety of genetically encoded, fluorescently tagged markers in hemocytes to follow their developmental dispersal throughout the embryo. Following collection of embryos at the desired stage of development, the outer chorion is removed and the embryos are then mounted in halocarbon oil between a hydrophobic, gas-permeable membrane and a glass coverslip for live imaging. In addition to gross migratory parameters such as speed and directionality, higher resolution imaging coupled with the use of fluorescent reporters of F-actin and microtubules can provide more detailed information concerning the dynamics of these cytoskeletal components.
Evans
Iwan R
IR
Department of Biology and Biochemistry, University of Bath.
Zanet
Jennifer
J
Wood
Will
W
Stramer
Brian M
BM
D000818:Animals; D002465:Cell Movement / Q000502:physiology*; D004331:Drosophila melanogaster / Q000166:cytology / Q000196:embryology*; D005456:Fluorescent Dyes / Q000737:chemistry; D049452:Green Fluorescent Proteins / Q000737:chemistry; D006434:Hemocytes / Q000737:chemistry / Q000166:cytology*
D005456:Fluorescent Dyes; D049452:Green Fluorescent Proteins