The Chen Liangyi team of the Institute of Molecular Medicine of Peking University and the Tanshan team of Huazhong University of Science and Technology invented an ultra-sensitive structural light ultra-high resolution microscope, Hessian SIM. This achievement was recently published online in full text on Nature Biotechnology, entitled "Fast, long-term, super-resolution imaging with Hessian structured illumination microscopy".
With 188 ultra-high resolution images per second, the Heisen structured light microscope has a spatial resolution of 85 nanometers and is capable of distinguishing 1/600 to 1/800 size of a single hair, while requiring less illuminance than commonly used. Confocal microscopy illumination is three orders of magnitude. Thanks to extremely low photobleaching and phototoxicity, 180,000 ultra-high resolution images are obtained by continuous sampling for 10 minutes at 100 Hz ultra-high resolution imaging, or ultra-high resolution imaging for 1 hour at 1 Hz ultra-high resolution imaging. Basically no photobleaching.
Compared with the stimulated radiation loss ultra-high resolution microscope (STED), which received the 2014 Nobel Prize in Chemistry, Heisen structured light microscopy has ultra-high resolution of living cells with extremely high temporal resolution and extremely low phototoxicity. Rate imaging has a significant advantage. For example, in the process of observing the fusion of intracellular vesicles with the cytoplasmic membrane to release neurotransmitters and hormones, both Heisen structured light microscopy and STED microscopy can observe the pores formed by vesicle fusion; however, the Heisen structural light microscope also resolves The vesicles fuse in four different intermediate states, including the vesicle opening 3 nanometer pores, vesicle collapse, fusion pore maintenance and the final fusion of the vesicles with the cytoplasmic membrane, truly visualizing the entire process of membrane pore formation (Figure 1 ).
On the one hand, this breakthrough is based on the new polarization rotating glass array designed by hardware, high-precision timing control program and the application of high numerical aperture objective lens; on the other hand, the innovative reconstruction algorithm draws on the human eye to distinguish between signal and noise. The mechanism proposes for the first time that biological samples are continuous in multidimensional space and time, while the noise is a completely random distribution of prior knowledge used to construct the Hessian matrix, guiding the reconstruction of ultra-high resolution fluorescence images.
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