In this protocol, we discuss general techniques for tracking the three-dimensional (3D) locations of the mosquito body, wings, legs, or other features of interest using videos. Tracking data must be acquired to produce detailed kinematics of moving mosquitoes. The software of focus for this protocol, DLTdv, was chosen for its widespread use and excellent support and because it is open-source. In addition, DLTdv allows both manual and automatic tracking. The automatic tracking can be done using a classic machine vision or machine-learning algorithm. The software supports both single-camera analysis and multicamera systems and can take advantage of sophisticated calibration algorithms, both for intrinsic lens distortion correction and for 3D DLT-based reconstruction. For this protocol, we assume all kinematic data is acquired post hoc through video analysis.
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