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-electro-mechanical 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 microm of tissue. The results also show that the isoplanatic half-width is approximately 19 microm resulting in a corrected field of view 38 microm in diameter around the guide-star.
Azucena
Oscar
O
Jack Baskin School of Engineering, Univ of California, Santa Cruz, 1156 High St, Santa Cruz, CA 95064, USA. azucena@soe.ucsc.edu
Crest
Justin
J
Cao
Jian
J
Sullivan
William
W
Kner
Peter
P
Gavel
Donald
D
Dillon
Daren
D
Olivier
Scot
S
Kubby
Joel
J
D000818:Animals; D003315:Cornea / Q000196:embryology*; D004330:Drosophila / Q000196:embryology; D004625:Embryo, Nonmammalian / Q000166:cytology; D005453:Fluorescence; D008863:Microspheres; D012029:Refraction, Ocular; D012030:Refractive Errors / Q000175:diagnosis*