Drosophila third-instar larval blood cells pumped anteriorly through the dorsal vessel. Live visualization of blood cells pumped within the dorsal vessel (heart) of a Nrg-GFP;PxnYFP larva. Blood cells enter the dorsal vessel at the posterior (bottom) and are rapidly pumped to the anterior. Red circles (first frame) indicate locations where pumped blood cells will appear in subsequent frames. Because individual cells were visible in consecutive frames of this movie, it allowed quantitation of the speed of the cells through the dorsal vessel. Red arrows in successive later frames point to the cells of interest. Time between frames, 0.325s. Total duration, 2.0 s. Fig. 1D was generated from individual frames of this movie. Movie is Movie S6 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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