Random access multiphoton (RAMP) microscopy for investigation of cerebral blood flow regulation mechanisms.

The processes by which blood flow is regulated at the capillary network level in the brain has been a source of continual debate. It is generally accepted that cerebral blood flow regulation occurs primarily at the arteriolar level. It has been recently suggested, however, that the capillary network is likewise under dynamic regulation. The exact mechanisms of capillary regulation remain unknown. Previously, the limiting factor in determining how the cerebrovascular network is regulated has been the speed at which multiphoton images of large (~200μm2) capillary and arteriole systems can be acquired. Conventional laser scanning microscopy systems are temporally limited in two dimensions. We have developed a Random Access Multiphoton (RAMP) microscope, which operates on the principles of Acousto-optic beam scanning and therefore has no moving parts, specifically for the purpose of imaging blood flow virtually simultaneously throughout the capillary network. We demonstrate the ability to survey blood flow simultaneously in 100 capillaries.