Three-dimensional data acquisition with aberrations correction for video-rate microscopy

摘要

We demonstrate a multimodal, multifocal, differential nonlinear optical microscope, which is equipped with a pair of deformable mirrors and a Shack-Hartmann sensor for dynamic wavefront manipulation. The optical wavefronts of a home built Yb:KGW femtosecond (1028 nm) laser-beams are engineered to perform multidepth focusing in differential mode with simultaneous corrections for optical aberrations. The 39-actuator deformable mirrors provide fast reshaping of the wavefront and optical aberrations correction of the diffraction-limited focal volume allowing for fast axial scanning. Combination of ~200 frames per second lateral scanning with fast refocusing enables a three-dimensional video rate scanning capability, which is essential for studying rapid dynamics in biological organisms, such as blood flow, cardiac contractions, and motility of microorganisms in a three-dimensional volume.