When using a dual scan path microscope for photostimulation it is often desirable to image and identify cells for stimulation with one scan path, then stimulate the cells using the other scan path. For this targeting to be accurate, alignment between the scanners is critical.
When using the photostimulation with a single scan path microscope, alignment is not necessary. If you are using a resonant-galvo-galvo scanhead however, it is important to have accurate settings for scan mirror angular ranges if you intend to use images collected in resonant mode to identify ROIs to stimulate. Details on how to do this can be found in the aspect ratio guide.
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Dual Scanhead setup for ScanImage. Two independent scan heads allow for simultaneous imaging and photostimulation. A dichroic mirror combines the two scan paths into the same objective.