Fast image reconstruction for real-time super

JSFR-SIM allows real-time observation of fine subcellular dynamics.

Image: JSFR-SIM allows real-time observation of fine subcellular dynamics.
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Credit: Zhaojun Wang and Ming Lei, Xi’an Jiaotong University

Super-resolution structured illumination microscopy (SR-SIM) is an excellent method for visualizing subcellular dynamics in living cells. SR-SIM can achieve fast, optically segmented (OS), super-resolution (SR) observation with hundreds to thousands of time points, the number of super-resolution frames for continuous imaging.

However, the reconstruction algorithm for OS-SR-SIM imposes a significant computational burden due to a complex workflow and large number of calculations. This effectively negates real-time imaging, as it takes 4 to 8 seconds to reconstruct a single SR image of 1024×1024 pixels. Microscope operators must first use the widefield mode to navigate the promising field of view for their examination, then switch to the SR-SIM mode to obtain the raw images for SR-SIM, then wait a long time to observe a single SR image , after special post-processing. The disjointed workflow of SIM configuration hinders the widespread adoption of SR-SIM among biologists.

Currently, parallel computing tools such as GPUs can be used to speed up SR reconstruction, but the result is limited to a modest raw image size.

As reported in Advanced Photonics, researchers from Xi’an Jiaotong University (XJTU, China) recently developed a rapid reconstruction algorithm, “Joint Space and Frequency Reconstruction” (JSFR-SIM), to address both the reconstruction speed and the limitation known as the problem with the missing cone. They combine a spatial domain processing technique with OS-SR-SIM implemented in the frequency domain, achieving an improved image reconstruction speed plus suppression of the blurred background in images of thick cells.

The reconstruction execution speed is 80 times faster than the widely used Wiener SIM. It is critical that the speed increase is not at the expense of image quality. To demonstrate the method’s performance, they performed real-time live cell observation of COS-7 cells, revealing insights into microtubule dynamics and rapid mitochondrial tubulation.

The JSFR-SIM method makes locating and imaging a super-resolution field of view a simple and routine process, allowing complex intracellular dynamics to be visualized in real time and resulting images to be quantified. It is compatible with different SIM modalities and shares the same hardware configuration as the conventional 2D SIM, so it can be easily applied to many new SIM modalities, including AO-SIM, pSIM and GI-SIM, to eliminate the computational burden , even when imaging thicker samples. This breakthrough promises to improve the work efficiency of biologists and facilitate SR-SIM as a routine tool in biomedical laboratories.

Read the open access article by Z. Wang et al., “High-speed image reconstruction for optically segmented, super-resolution structured illumination microscopy”, Adv. photon. 4(2) 026003 (2022) doi 10.1117/1.AP.4.2.026003.

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