Acquisition of High-Quality Digital Video of Drosophila Larval and Adult Behaviors from a Lateral Perspective

Current modes of scientific publishing support the submission of digital video media files as supplemental material to a manuscript. These video media files are often used to convey qualitative information to the reader that is easier observed than described in text or by a single photograph. Here we outline a simple methodology for acquiring high quality digital video of drosophila mutant phenotypes from a lateral perspective, Although This protocol is easily adaptable to virtually any stereo microscope system coupled to a digital camera, with the capability of acquiring video, details are provided on the system we use in our lab.

We acquire our video using an Olympus SZ 61 ocular stereo microscope, coupled to a commercial Canon G 10 digital camera. The one half XC mount of the photo tube port of the stereo microscope was reconfigured with a one XC mount. A digital camera coupler was mounted to the one XC mount.

Two step-down rings 58 millimeter to 48 millimeter and 48 millimeter to 43 millimeter were mounted to the camera coupler. An adapter kit for a Canon G 10 camera was mounted. Bridging the connection to the Canon G 10 camera To image.

Third In star larvae from the lateral perspective, tape a permanent marker to the stage of the stereo microscope. The Marker cap should occupy approximately one quarter of the vertical field observed in the camera LCD monitor. We use markers as an alternative stage because they come in an assortment of colors that allow us to color code and differentiate the genotypes of larva being Imaged On the marker cap.

Mark the left and right boundaries Of the field of view observed in the camera LCD monitor. Select a third instar larva to image. Ensure the larva is clean by washing it thoroughly in water.

Focus the microscope on the edge Of the marker cap and begin acquiring digital video. Place the larvae on the side of the marker cap just outside the field of view. Placement of the larvae on the side of the marker cap allows the camera to record movement of the larvae From a lateral perspective, it helps to keep the larvae moist with water so they don't fall off the side of the marker cap.

However, be careful not to use too much water. Gently poke and prod the larvae With a paintbrush to coerce it to crawl across the field of view. Be patient.

The larva rarely cooperate and often have to be returned to the starting point many times until they crawl straight across the field. We usually record 10 to 15 minutes of uninterrupted digital video footage and crop and remove all unnecessary footage. Post-acquisition with digital video editing Software to image adult Ultra sophala.

From the lateral perspective, we use disposable 1.5 milliliter spectroscopic polystyrene cuvettes as cages to contain the flies. We prefer the smaller volume qve because the reduced Volume capacity restricts the movement of the fly. Place the adult Drosophila in the qve and plug the end with a small cotton ball.

Ensure the cotton ball is packed tight enough to occupy the large cap space. Place the Qve on the microscope stage and properly align with the LCD monitor on the camera. Focus the microscope and begin acquiring digital video.

We usually record 30 to 45 minutes of uninterrupted digital movie footage and crop and remove all unnecessary footage post acquisition with digital video editing Software. We have Successfully used this technique to visualize Drosophila third instar larvae with abnormal crawling phenotypes. We have also used this technique to visualize adult grooming behaviors as well as adult mutant phenotypes.

We have just Demonstrated a very simple and accessible technique for acquiring high quality digital video of drosophila larvae in adults. The benefit of this approach is that video is acquired from a lateral perspective, allowing direct observation and analysis of aberrant locomotive behaviors from the more informative side view. Thanks for watching and good luck with your experiments.