Multiphoton imaging microscopy at deeper layers with adaptive optics control of spherical aberration

Abstract

Despite the inherent confocality and optical sectioning capabilities of multiphoton microscopy, threedimensional (3-D) imaging of thick samples is limited by the specimen-induced aberrations. The combination of immersion objectives and sensorless adaptive optics (AO) techniques has been suggested to overcome this difficulty. However, a complex plane-by-plane correction of aberrations is required, and its performance depends on a set of image-based merit functions. We propose here an alternative approach to increase penetration depth in 3-D multiphoton microscopy imaging.