Light Spot Assay: A Method to Investigate Drosophila Phototactic Behavior
DEŞİFRE METNİ
– Begin by selecting a third instar larva, the largest of the larval forms, from the culture medium, and wash it with water için remove any food. Since the larval eyes, or Bolwig organs, are located videodan the upper front of the larva, place the larva videodan the center of an agar plate with the light source above the animal. Turn off the room light and allow the larva için acclimate için the dark.
Since the larva is barely detectable videodan a light spot made by visible light, turn on infrared LED lights için illuminate the animal for the camera. Wait until the animal begins crawling straight için control for its path. And then rotate the plate so that the larva crawls directly toward the light spot. Record the animal crawling into the light and then away from the light spot. This light-avoidance tactic helps larvae avoid predators by burrowing into food.
In the following example, we will perform the light spot assay için test the ability of different genetic strains of Drosophila larvae için escape light.
– Before introducing the larva için the imaging setup, acquire a short video of the light position, and İsim the unfiltered control background video “lightarea1”. Place the filter back on the camera lens, and turn on a light far from the experimental device at as low a setting as possible while still allowing the larvae için be clearly observed. Use a spoon için remove some larvae from the culture medium, and gently select a third instar larva.
Wash the larva videodan distilled water, and place the larva videodan the center of the agar plate. Gently remove any excess water from the larva, and turn off the room light. Allow the larva için acclimate for two minutes videodan the dark environment before turning on the infrared LED light. The following light-avoidance behavior experiment needs için be carried out videodan the dark. For the convenience of demonstration, we filmed videodan the bright light.
Gently brush the larva için the center of the plate. When the larva begins için crawl straight, rotate the plate için make the larva head towards the light spot. Click Capture, Setup, and Video Capture için select the save path. And click Start Recording için record. Allow the lava için crawl toward, enter, and leave the light spot until it is nearly out of the field of view, then click Stop Recording.
Move the filter away from the camera, and acquire a short video of the position of the light spot. Then İsim the post as a unfiltered video, “lightarea2”, and compare this video with the “lightarea1” video için ensure that the light spot position is not changed.
