Rueckel Markus, Mack-Bucher Julia A, Denk Winfried
Department of Biomedical Optics, Max-Planck Institute for Medical Research, Jahnstrasse 29, D-69120 Heidelberg, Germany.
Proc Natl Acad Sci U S A. 2006 Nov 14;103(46):17137-42. doi: 10.1073/pnas.0604791103. Epub 2006 Nov 6.
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.
双光子显微镜的图像质量常常会因样本引起的波前像差而下降。我们在此证明,通过基于相干选通背向散射光的波前传感(相干选通波前传感,CGWS)的自适应波前校正以及由可变形镜进行的波前控制,即使在活的生物样本中,双光子显微镜的分辨率和信号大小也能得到显著提高。即使对于强像差,也能恢复接近衍射极限的焦点。基于CGWS的波前校正应适用于具有广泛散射特性的样本,并且即使在快速扫描速度下也应能够进行实时逐像素校正。