Adaptive Wavefront Correction in Two-photon Microscopy - Coherence-Gated Wavefront Sensing

The focus of a two-photon microscope is often degraded by inhomogeneities in the refractive index within biological specimens. In this book it is shown for various specimens, even for living zebrafish, that the resolution and the fluorescence signal of a two-photon microscope can be substantially improved by using adaptive optics, i.e. wavefront correction based on coherence-gated wavefront sensing (CGWS). The advantage of using CGWS relies on the fact that the wavefront distortions are sensed by backscattered instead of fluorescent light. Thus, neither photodamage nor photobleaching occurs and wavefront distortions can be sensed in less than 1 microsecond with an accuracy of lambda/50, even in strongly scattering samples. Furthermore, CGWS is thoroughly investigated for all relevant parameters affecting the measurement process, such as coherence length, polarization of the light, density of scatterers, and coherence-gate position. The book is mainly addressed to experts in the field of optical physics and light microscopy.