KEYWORDS: Adaptive optics, Super resolution microscopy, Microscopes, Super resolution, 3D image processing, Stereoscopy, Optical aberrations, Microscopy, Luminescence, Imaging systems
Super-resolution microscopy allows the observation of sub-cellular structures with a resolution beyond diffraction limit of conventional fluorescence microscopy. However, most super-resolution microscopes have a limited imaging depth due to the inhomogeneous refractive index of the sample that leads to optical aberrations. Adaptive optics has been successfully adopted by many imaging techniques, including 3D Structured illumination microscopy (SIM). We use a fast deformable mirror to modulate the wavefront of fluorescence to compensate for optical aberration and changing focus position at the same time. Adaptive optics successfully extends the depth, the range and the speed of 3D-SIM imaging.
Specimen induced aberrations can have detrimental effects in all types of high-resolution microscope. In this study, we present a sensorless technique that uses a deformable mirror (DM) to correct aberrations of both the system and sample. Using a laser-free confocal microscope, with patterned disk illumination and detection. The system is based on a commercial confocal module (Clarity, Aurox Ltd., UK) that uses Light Emitting Diode (LED) illumination to obtain optically sectioned 3D images. The results obtained show that the setup was able to correct aberrations of biological samples used in the study. These systems will help researchers working on various biological systems to obtain improved quality images when focussing deep into thick specimens.
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