Adaptive wavefront correction in two-photon microscopy using coherence-gated wavefront sensing.
The image quality of a two-photon microscope is often degraded by wavefront aberrations induced by the specimen. We demonstrate here that resolution and signal size in two-photon microcopy can be substantially improved, even in living biological specimens, by adaptive wavefront correction based on sensing the wavefront of coherence-gated backscattered light (coherence-gated wavefront sensing, CGWS) and wavefront control by a deformable mirror. A nearly diffraction-limited focus can be restored even for strong aberrations. CGWS-based wavefront correction should be applicable to samples with a wide range of scattering properties and it should be possible to perform real-time pixel-by-pixel correction even at fast scan speeds.
Rueckel
Markus
M
Department of Biomedical Optics, Max-Planck Institute for Medical Research, Jahnstrasse 29, D-69120 Heidelberg, Germany. markus.rueckel@mpimf-heidelberg.mpg.de
Mack-Bucher
Julia A
JA
Denk
Winfried
W
D000818:Animals; D017930:Genes, Reporter / Q000235:genetics; D007814:Larva; D036641:Microscopy, Fluorescence, Multiphoton / Q000295:instrumentation* / Q000379:methods*; D015027:Zebrafish