Background: In respiratory gated radiotherapy, low latency between target motion into and out of the gating window and actual beam-on and beamoff is crucial for the treatment accuracy. However, there is presently a lack of guidelines and accurate methods for gating latency measurements. Purpose: To develop a simple and reliable method for gating latency measurements that work across different radiotherapy platforms. Methods: Gating latencies were measured at a Varian ProBeam (protons, RPM gating system) and TrueBeam (photons, TrueBeam gating system) accelerator. A motion-stage performed 1 cm vertical sinusoidal motion of a marker block that was optically tracked by the gating system. An amplitude gating window was set to cover the posterior half of the motion (0-0.5 cm). Gated beams were delivered to a 5 mm cubic scintillating ZnSe:O crystal that emitted visible light when irradiated, thereby directly showing when the beam was on. During gated beam delivery, a video camera acquired images at 120 Hz of the moving marker block and light-emitting crystal. After treatment, the block position and crystal light intensity were determined in all video frames. Two methods were used to determine the gate-on (τ on ) and gate-off (τ off ) latencies. By method 1, the video was synchronized with gating log files by temporal alignment of the same block motion recorded in both the video and the log files. τ on was defined as the time from the block entered the gating window (from gating log files) to the actual beam-on as detected by the crystal light. Similarly, τ off was the time from the block exited the gating window to beam-off. By method 2, τ on and τ off were found from the videos alone using motion of different sine periods (1-10 s). In each video, a sinusoidal fit of the block motion provided the times T min of the lowest block position. The mid-time, T mid-light , of each beam-on period was determined as the time halfway between crystal light signal start and end. It can be shown that the directly measurable quantity T mid-light − T min = (τ off +τ on )/2, which provided the sum (τ off +τ on ) of the two latencies. It can also be shown that the beam-on (i.e., crystal light) duration ΔT light increases linearly with the sine period and depends on τ off − τ on : ΔT light = constant•period+(τ off − τ on ). Hence, a linear fit of ΔT light as a function of the period provided the difference of the two latencies. From the sum (τ off +τ on ) and difference (τ off − τ on ), the individual latencies were determined. Results: Method 1 resulted in mean (±SD) latencies of τ on = 255 ± 33 ms, τ off = 82 ± 15 ms for the ProBeam and τ on = 84 ± 13 ms,τ off = 44 ± 11 ms for the This is an open access article under the terms of the Creative Commons Attribution-NonCommercial License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited and is not used for commercial purposes.