Adaptive optics wide-field microscopy using direct wavefront sensing

Abstract

We report a technique for measuring and correcting the wavefront aberrations introduced by a biological sample using a Shack-Hartmann wavefront sensor, a fluorescent reference source, and a deformable mirror. The reference source and sample fluorescence are at different wavelengths to separate wavefront measurement and sample imaging. The measurement and correction at one wavelength improves the resolving power at a different wavelength, enabling the structure of the sample to be resolved.

Optical Property Analyses of Plant Cells for Adaptive Optics Microscopy

“…[6,7] AO microscopy has successfully been used to cancel the disturbances caused by cellular structures in wide-field microscopy, [8] laser-scanning confocal microscopy, [9,10] multi-photon microscopy, [11][12][13] harmonic generation microscopy, [14,15] and superresolution stimulated emission depletion microscopy, [16,17] with or without the wavefront sensor. [18] So far, most AO microscopy has observed artificial samples and animal tissues; plant cells and tissues have not been examined by AO microscopy yet.…”

Strategies to overcome photobleaching in algorithm-based adaptive optics for nonlinearin-vivoimaging

“…The reference sources employed were either injected fluorescent microspheres or nonlinear guide stars created by the TPEF. [14][15][16] A drawback of those closed loop methods is the loss of power due to the continuous monitoring of the wavefront and/or use of a spatial light modulator as well as the complexity of the setup. Additionally, measuring distorted wavefronts in microscopy is not trivial and generally requires a point source emitter in the sample.…”

Light sheet adaptive optics microscope for 3D live imaging

“…In [48,42], fluorescent beads have been used respectively on a widefield and confocal microscope. The bead supplies a point-like source for the wavefront sensor.…”

“…This works extremely well, but is relatively slow and cannot easily be used on dynamic samples. More recent work has been published on full closed loop AO [48,61,42,50,51]. A major challenge for AO microscopy is obtaining a good signal for the WFS.…”