Wavefront aberration measurements and corrections through thick tissue using fluorescent microsphere reference beacons

We present a new method to directly measure and correct the aberrations introduced when imaging through thick biological tissue. A Shack-Hartmann wavefront sensor is used to directly measure the wavefront error induced by a Drosophila embryo. The wavefront measurements are taken by seeding the embryo with fluorescent microspheres used as artificial guide-stars. The wavefront error is corrected in ten millisecond steps by applying the inverse to the wavefront error on a micro-electromechanical deformable mirror in the image path of the microscope. The results show that this new approach is capable of improving the Strehl ratio by 2 times on average and as high as 10 times when imaging through 100 mu m of tissue. The results also show that the isoplanatic half-width is approximately 19 mu m resulting in a corrected field of view 38 mu m in diameter around the guide-star. (C) 2010 Optical Society of America