Implementation of a Shack-Hartmann wavefront sensor for the measurement of embryo-induced aberrations using fluorescent microscopy

Adaptive optics (AO) improves the quality of astronomical imaging systems by using real time measurement of the turbulent medium in the optical path using a guide star (natural or artificial) as a point source reference beacon. AO has also been applied to vision science to improve the current view of the human eye. This paper will address our current research focused on the improvement of fluorescent microscopy for biological imaging utilizing current AO technology. A Shack-Hartmann wavefront sensor (SHWS) was used to measure the aberration introduced by a Drosophila Melanogaster embryo with an implanted 1 micron fluorescent bead that serves as a point source reference beacon. The measurements show an average peak-to-valley and root-mean-square (RMS) wavefront error of 0.77 micrometers and 0.15 micrometers, respectively. The Zernike coefficients have been measured for these aberrations which indicate that the correction of the first 14 Zernike coefficients should be sufficient to correct the aberrations we have obtained. These results support the utilization of SHWS for biological imaging applications and that a MEMS deformable mirror with 1 micron of stroke and 100 actuators will be sufficient to correct these aberrations. The design, assembly and initial results for the use of a MEMS deformable mirror, SHWS and implanted fluorescent reference beacon for wavefront correction will also be discussed.