High dynamic range structured illumination microscopy based on per-pixel coding

Abstract

Structured illumination microscopy (SIM) can achieve optical sectioning with high resolution, and have aroused extensive research interest. In SIM, a set of high-contrast illumination patterns are projected onto the sample to modulate the surface height information, and then, a decoding algorithm is applied to the modulated pattern images for high-quality optical sectioning. Applied to samples with large dynamic range of reflectivity, however, SIM may fail to achieve high quality sectioning for accurate surface reconstruction. Herein, an active digital micromirror device (DMD) based illumination method using per-pixel coded strategy is proposed in SIM to realize high-quality measurement for surface with complex reflection characteristics. In this method, the mapping relationship between DMD and the camera is established pixels by pixels, which enables the illumination intensity on the sample surface can be flexibly modulated by DMD pixel-level modulation corresponding to reflectivity distribution of the surface, and allows the camera pixels always to have reasonable exposure intensity for high precision measurement. More importantly, we put forward an adaptive light intensity control algorithm to improves the signal-to-noise ratio of acquired images without compromising modulation depth of pattern and measurement efficiency. Extensive comparative experiments were conducted and demonstrated that the proposed method can retrieve the surface morphology information of micro-scale complex reflectivity surfaces with high accuracy.