Transient release of a cluster of tissue-bound blood cells into circulation in a Drosophila third-instar larva. Real-time movie of blood cells visualized from dorsal side of a Nrg-GFP;PxnYFP larva. A sessile cluster of blood cells breaks apart and some of the cluster enters circulation and flows posteriorly and laterally. However, 4 s later, the released cells reattach to a nearby tissue (tracheal dorsal trunk). Red arrow in frame 1 shows the trajectory of the released cells. Time between frames, 0.325 s. Total duration, 8 s. Fig. 1H was generated from individual frames of this movie. Movie is Movie S7 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 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 real-time recording, frames were captured every ≈400 ms over a 4- to 40-s 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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