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Abstract
High-speed volumetric imaging technique is a useful tool to record and analyze biological activity. However, current volumetric imaging methods suffer from low imaging speed, poor contrast or resolution. Point-scanning microscopy and light-sheet fluorescence microscopy (LSFM) can effectively suppress the background noise, but their mechanical scanning strategy limits the speed of volumetric imaging. Light-field microscopy (LFM) captures 3D information without any scanning device, but constrained by the whole-volume illumination and the fixed microlens array, the contrast is low and the resolution is fixed and impossible to be optimized for different imaging scenarios. Here we report a selective volume microscopy (SVM) based on programmable light field microscopy (PLFM) with an engineered point spread function (PSF) strategy, which achieves flexible resolution tuning combined with selective illumination for optimized spatiotemporal resolution. By selectively illuminating and programmably detecting the volumes of interest (VOI) of the sample, SVM improved reconstruction contrast and spatial resolution by ∼2.6 times and ∼1.2 times in comparison with PLFM. The improvement of reconstruction quality of SVM is demonstrated by imaging the fluorescent beads and larval zebrafish across 665 μm × 665 μm × 200 μm at 100 Hz.
