Fast reconstruction algorithm for structured illumination microscopy

Abstract

Structured illumination microscopy (SIM) is a powerful technique for providing super-resolution imaging, but its reconstruction algorithm, i.e., linear reconstruction structured illumination microscopy (LRSIM) algorithm in the Fourier domain, limits the imaging speed due to its computational effort. Here, we present a novel reconstruction algorithm that can directly process SIM data in the spatial domain. Compared to LRSIM, this approach uses the same number of frames to achieve a comparable resolution but with a much faster processing speed. Our algorithm was verified on both simulated and experimental data using sinusoidal pattern illumination. Moreover, this algorithm is also applicable for speckle pattern illumination.

Recent Progress in the Correlative Structured Illumination Microscopy

“…In SIM, the sinusoidal excitation light is superimposed on the sample, and when this is convolved through the objective lens, the observed fluorescence emission takes the form of a Moiré fringe pattern. This pattern represents a mix of frequencies from both the excitation light and the object, with frequency information higher than the cutoff frequency [38,39].…”

Achieving Increased Resolution and Reconstructed Image Quality with Intensity and Gradient Variance Reweighted Radial Fluctuations

“…The above-mentioned super-resolution techniques have established themselves as important parts of cell biology research because of their respective advantages in spatial resolution, temporal resolution, and system complexity. For example, the temporal resolutions of SIM and STED are higher than STORM/PALM so they are conducive to dynamic live-cell imaging. − However, their spatial resolutions are relatively inferior to those of STORM/PALM. In addition, SIM and STED require stable and precise optical systems and complex setups, which unavoidably improve the difficulty and complexity of system construction.…”

Electron-beam patterned calibration structures for structured illumination microscopy

“…In SIM, data is reconstructed from a stack of images recorded after modulation with mutually rotated and shifted inhomogeneous illumination patterns. Data reconstruction is computationally intensive and various SIM reconstruction algorithms have been reported over the past years 1 – 8 . These algorithms involve parameters, for instance for regularization and apodization filters, that are generally adjusted at will by the user.…”