Adhesive capture of a sheet of blood cells at the wound site in a Drosophila third-instar larva. Time-lapse movie of blood cells visualized from the dorsal side of a Nrg-GFP;PxnYFP larva. Blood cells accumulate at the larval wound site (area outlined in white in first and last frames) over a span of 4 h. Time between frames, 5 min. Individual blood cells and small clusters gradually bind to the left side of the wound site over the first two hours, followed by capture of a large sheet of cells within a 10-min interval that fills almost the entire right side of the wound area. Fig. 3C was generated from individual frames of this movie. Movie is Movie S11 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.
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.
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