| OCR Text |
Show 83 3.3.12 Whole mount in situ hybridization: Drosophila Drosophila whole mount in situ hybridization was performed as described previously (Byars et al., 1999). 3.3.13 Body length: zebrafish Zebrafish from a single home tank were anesthetized using Tricaine in shallow water. Images were acquired of immobilized, nonoverlapping fish with a ruler for scale. Body length was calculated by measuring the distance between the mouth and the anterior edge of the tail fin, using ImageJ. 3.3.14 Novel tank diving test 5 fish from lef1+/- incrosses were raised per tank starting at 5 dpf. lef1 mutants and controls were separated at 15 dpf. Novel tank diving tests were performed on 16 dpf larvae during the early afternoon of the same days, before lef1 mutants start to display surfacing behavior at 20 dpf. Novel rectangular tanks (16.6 cm × 9.5 cm × 12.3 cm) were illuminated by a centered white light, and videos were acquired with a mounted Nokia Lumia 640 phone 1080p camera. For each experiment, single mutant and control larvae were netted and then removed simultaneously from their home cages and transferred to novel tanks with identical water volume. Any larvae that were not immediately released from the net were excluded from analysis. The order of netting mutant and control fish was rotated between trials. Videos were then imported and analyzed using Ethovision XT v.11.5 (Noldus, Leesburg, VA), with a tracking period of 2 min beginning 1 min after release into |
