Adaptive optics confocal microscopy using fluorescent protein guidestars for brain tissue imaging

摘要

Optical aberrations due to the inhomogeneous refractive index of tissue degrade the resolution and brightness of images in deep tissue imaging. We introduce a direct wavefront sensing method using cellular structures labeled with fluorescent proteins in tissues as guide-stars. As a non-invasive and high-speed method, it generalizes the direct wavefront sensing method for adaptive optics microscopy. An adaptive optics confocal microscope using this method is demonstrated for imaging of mouse brain tissue. The confocal images with and without correction are collected. The results show increased image contrast and 3X improvement in the signal intensity for fixed mouse tissues at a depth of 70 μm. The images of the dendrite and spines are much clearer after correction with improved contrast. The Strehl ratio is improved from 0.29 to 0.96, a significant 3.3X improvement.