Interlaced two-photon volumetric scanning for measurement of capillary network dynamics

We combined a home-built two photon microscope with a Bessel focus capable of volumetric imaging with a custom scan pattern that allowed us to generate image pairs with a time delay of ~12 ms. This approach allows us to track red blood cell (RBC) flow along individual capillaries and therefore calculate RBC speed. We demonstrate the ability to measure flow speeds in capillaries across a 475x475x150 µm volume which will enable us to monitor network wide changes in capillary flow.