Chinese and foreign scientists collaborate to develop a new super-resolution optical microscope imaging system
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[China Instrument Network Instrument Development] Recently, Zheng Wei, a researcher at the Shenzhen Institute of Advanced Technology of the Chinese Academy of Sciences, and Hari Shroff, a professor at the National Institutes of Health, have successfully developed a new two-photon excitation super-resolution optical microscopy imaging system.

The left picture shows the contrast between the results of the two-photon imaging (2P WF), two-photon super-resolution imaging (2P ISIM) and two-photon super-resolution (2P ISIM AO) combined with adaptive optics for the Drosophila brain slices. In contrast to the three-dimensional imaging of cells located 150 microns deep in the collagen gel, it can be seen that the resolution of the new technology has improved significantly both horizontally and vertically. The bottom right panel shows three-dimensional observations of an hour continuously during the embryonic development of the nematode. The normal cell division process proves that this technique can be used for dynamic research of embryonic development.
The system has both super-resolution optical microscopy imaging capabilities and large-depth 3D imaging capabilities, enabling the advancement of optical super-resolution imaging to a record-breaking 250 micron depth, corresponding research results. Resolution microscopy imaging was recently published in Nature Methods.
“Seeing fine” and “seeing it deeply” are two major challenges faced by the field of optical microscopy imaging. After decades of unremitting efforts by scientific researchers, whether in “seeing fine” or “deeply seeing”, A number of innovative technologies have emerged and achieved great success. However, optical microscopy imaging technology with both "see fine" and "deep view" functions is rare.
In this study, Zheng Wei et al. used Two-Photon Microscopy (TPM) with deep biological tissue imaging capabilities and Instant Structured Illumination Microscopy with super-resolution imaging capabilities. , ISIM) Combines organically to achieve two-photon excitation of super-resolution microscopy imaging. At the same time, the researchers successfully used adaptive optics (AO) technology to overcome the wavefront phase distortion problem caused by biological tissue, and ultimately achieved a lateral resolution of 176 nm, a vertical resolution of 729 nm, and a detection of 250 μm. Depth imaging effect. Using this technology, high-resolution 3D imaging studies can be performed on cells, nematode embryos and larvae, fruit fly brain slices, and zebrafish embryos, and imaging results are significantly better than conventional two-photon imaging. It is worth mentioning that since this technology improves the efficiency of photon utilization and thus reduces the required laser power, it can perform long-time, high-definition three-dimensional dynamic observations of the development process of nematode embryos. In the process of continuous three-dimensional imaging for up to one hour, it did not have any effect on the development of nematode embryos. This technique plays an important role in embryonic development research.
The research was supported by the National Natural Science Foundation of China, the National Key Basic Research and Development (“973”) Program, and Shenzhen Overseas High-level Talent Innovation and Peacock Program.
(Original Title: Shenzhen Advanced Institute, etc. Progress in Super Resolution Optical Microscopy Imaging)

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