Description

Adhesive capture of individual blood cells at a Drosophila third-instar larval wound site. Time-lapse movie of blood cells visualized from the dorsal side of a Nrg-GFP;PxnYFP larva. Individual blood cells adhere to the wound site (area outlined in white in first and last frames) over a period of 2 h. Time between frames, 5 min. Movie is Movie S8 of Proc Natl Acad Sci (2008). 105: 10017-10022. Movies from this publication include CIL #s 25607, 25608, 25609, 25610, 25611, 25612, 25613, 25614, 25615, 25616, 25617.

Technical Details

Third-instar [w, Nrg-GFP;Pxn-Gal4, UAS-2xeYFP] larvae were wounded and mounted dorsal side up on a glass slide with the anterior and posterior ends of the larvae taped down to varying degrees to prevent locomotion and constrain body peristalsis. Larvae were imaged on a Leica MZ16FA fluorescent stereomicroscope using a Planapo 1.0× objective, and images were captured on a monochrome Leica DFC350FX digital camera. For time-lapse imaging, frames were captured every 2 or 5 min over a 2- to 4-h period. Images were obtained by using the Scope-Pro Advanced Acquisition plug-in, and image analysis was carried out with Image-Pro AMS ver. 5.1 Software (Media Cybernetics). The object-tracking tool was used to manually track specific blood cells in a time series. Velocities of tracked cells were calculated by using coordinates provided by the software.