Abstract

Structured illumination microscopy (SIM) is a widely used super resolution imaging technique that can down-modulate a sample's high-frequency information into objective recordable frequencies to enhance the resolution below the diffraction limit. However, classical SIM image reconstruction methods often generate poor results under low illumination conditions, which are required for reducing photobleaching and phototoxicity in cell imaging experiments. Although denoising methods or auxiliary items improved SIM image reconstruction in low signal level situations, they still suffer from decreased reconstruction quality and significant background artifacts, inevitably limiting their practical applications. In order to improve the reconstruction quality, second-order optimized regularized SIM (sorSIM) is designed specifically for image reconstruction in low signal level situations. In sorSIM, a second-order regularization term is introduced to suppress noise effect, and the penalty factor in this term is selected to optimize the resolution enhancement and noise resistance. Compared to classical SIM image reconstruction algorithms as well as to those previously used in low illumination cases, the proposed sorSIM provides images with enhanced resolution and fewer background artifacts. Therefore, sorSIM can be a potential tool for high-quality and rapid super resolution imaging, especially for low signal images.

Highlights

  • In order to overcome diffraction limit in optical imaging, various super resolution imaging techniques have been proposed, including stimulated emission depletion microscopy (STED) [1,2], photoactivated localization microscopy (PALM)/stochastic optical reconstruction microscopy (STORM) [3,4], super resolution optical fluctuation imaging (SOFI) [5] and structured illumination microscopy (SIM) [6,7,8], etc

  • As the cost function of Fourier ptychography (FP)-SIM does not consider noise suppression, high-quality super resolution images can hardly be extracted from low signal to noise ratio (SNR) structured illuminated images; while second order regularization term is introduced in second-order optimized regularized SIM (sorSIM) to significantly reduce the noise influence on image reconstruction

  • We designed and evaluated a method that allows to rapidly reconstruct high-quality super resolution images from captured structured illuminated images especially relevant for low signal level situations. This is relevant for live cell imaging in order to reduce photobleaching and phototoxicity

Read more

Summary

Introduction

In order to overcome diffraction limit in optical imaging, various super resolution imaging techniques have been proposed, including stimulated emission depletion microscopy (STED) [1,2], photoactivated localization microscopy (PALM)/stochastic optical reconstruction microscopy (STORM) [3,4], super resolution optical fluctuation imaging (SOFI) [5] and structured illumination microscopy (SIM) [6,7,8], etc. Fourier ptychography (FP) [19,20,21,22,23] and RichardsonLucy deconvolution [24,25,26,27,28] based SIM image reconstruction methods have been proposed to pursue fast iteration convergence and rapid processing speed These methods provide extremely high accuracy in super resolution image reconstruction in rather high signal to noise ratio (SNR) conditions. In order to prolong observation periods especially for live cell imaging, illumination intensity is often decreased aiming at reducing photobleaching and phototoxicity, while inevitably decreasing the signal level Both classical and blind SIM image reconstruction methods cannot provide high quality results in low SNR conditions, and limiting their applications in long term live cell imaging. By balancing the trade-off between resolution enhancement and noise resistance, sorSIM may become a useful tool for long term, live cell super resolution imaging with low illumination conditions

Principle
Experiments
Findings
Conclusion
Full Text
Published version (Free)

Talk to us

Join us for a 30 min session where you can share your feedback and ask us any queries you have

Schedule a call