Confocal Microscopy (CM) enables 3-D high-resolution images acquisition of biological samples by confocal pinhole application. Whereas transverse scanning can be provided by implementation of scanning mirrors, axial scanning can be achieved by moving the sample, which limits the acquisition speed. On the other hand, with coherence gating Optical Coherence Microscopy (OCM) allows to generate micrometer resolution, cross-sectional images and volumetric data on the internal structure of back-scattering objects. The aim of our study was to assess imaging performance of different designs of CM and OCM with tunable lens technology. Mechanical scanning in axial direction was replaced by remote control of the objective focus position with the two types of lenses: Electrically Tunable Lens (ETL; up to 500 Hz) and Acousto-Optic Lens (AOL; 250 kHz). We compared focus tuning ability of ETL and AOL technologies by applying beam profiling and wavefront sensing. We assessed the impact of tunable lens technology on the CM and OCM system performance. We designed the confocal microscopic system with Bessel beam illumination to extend the depth-of-focus. The axial scanning of the collection point will be provided by our AOL. Combination of Bessel beam illumination and AOL allowed high-speed image acquisition from well-controlled depth positions along with better quality of PSF due to self-healing properties of the Bessel beam. We compared image quality of the proposed configurations with the standard design using biological specimens. To conclude, tunable lens technology implemented to CM and OCM instruments enables enhancement of instrument performance.
Krzysztof Szulzycki, Viktoriya Savaryn, and Ireneusz Grulkowski, "High-speed remote focusing for improvement of 3D imaging performance in confocal microscopy and optical coherence microscopy (Conference Presentation)," Proc. SPIE 10499, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXV, 1049910 (Presented at SPIE BiOS: January 31, 2018; Published: 14 March 2018); https://doi.org/10.1117/12.2290074.5751609925001.
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