Adaptive Optics in Multiphoton Microscopy

Abstract

Any type of microscopy faces the problem in an attenuated signal level and reduced optical resolution due to optical aberration. To overcome this problem, adaptive optics was implemented in a two-photon fluorescence microscope. Using a “sensorless” approach, this study corrected the aberration in the system using two different excitation colors. As methodology, the point spread function was compared before and after applying adaptive optics. This study demonstrated that adaptive optics improves the resolution of the microscope for both excitation wavelengths. The aberration correction was then monitored as a function of depth. The result showed an improvement in the optimization metric as imaging depth is increased. Thus, adaptive optics offers improved imaging of the sample at deeper depths with better optical resolution and higher signal-to-noise ratio.