ABSTRACT
Two-photon fluorescence microscopy, introduced in 1990, dramatically improved the ability to see inside strongly scattering biological tissues. The enhanced Signal to Noise Ratio and reduced tissue absorbance and scattering in two photon fluorescence microscopy allowed the maximal penetration depth into a brain tissue to increase from less than 100 µm up to 500 µm. Parallelization of the data acquisition process, in both the illumination and detection parts of the microscope, will enable significantly increased imaging speeds, generally up to the level of parallelization. Wide-field illumination offers a potential solution for fast volume imaging, but its major drawback is that it lacks any optical sectioning. Remote scanning is defined as an axial shift of the imaging system focal plane without moving the objective or the sample. Because of the challenges associated with mechanical scanning of the objective lens, remote scanning solutions can be very useful for rapid imaging systems and multi-modal microscopy systems.
