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As the life science research continues to deepen, scientists are increasingly demanding the use of optical microscopy equipment for research. Especially for biological living microscopic imaging, the scattering of light by biological tissues greatly increases the noise, which seriously affects the spatial resolution and imaging depth. In order to improve the imaging depth, two-photon excitation laser scanning fluorescence microscope technology has been widely used in neuroimaging and other fields since its introduction in the 1990s, but its point-by-point scanning imaging mode has severely restricted the imaging speed. Because the depth of field of the high-resolution optical microscope is very small, to complete the three-dimensional imaging of the sample, usually dozens of layers or even hundreds of two-dimensional images are obtained by stacking and reconstruction. The image acquisition and processing generally takes several minutes or even tens of minutes. It is very difficult to acquire and display 3D images in real time.
This time, the research team of the Chinese Academy of Sciences, based on the principle of binocular vision and the extended coke field generated by the Bessel beam, proposed a three-dimensional laser beam scanning device consisting of four galvanometers, realizing the lateral position and inclination of the Bessel beam. A total of three dimensions of control break through the limitation that traditional laser scanning with only two degrees of freedom cannot switch perspectives in real time. Through the optimization design and control of the four-way mirror three-dimensional scanning device, a three-degree-of-freedom scanning of the Bessel beam is realized, and the dual-angle switching can be performed in the order of milliseconds, thereby solving the problem in the laser scanning stereomicroscope imaging system. For the first time, a three-dimensional microscopic imaging and display system based on dual-view real-time laser scanning has been realized.
The system can perform stereoscopic dynamic imaging of the sample and real-time binocular stereoscopic observation. The three-dimensional imaging speed is improved by one to two orders of magnitude compared to the conventional point-by-point scanning method. The two-photon stereomicroscope system provides a new observation tool for three-dimensional real-time imaging and display of living organisms. According to the researchers, with this new microscopy device, users can watch the dynamic 3D microscopic world in real-time as if watching a stereoscopic movie, without the need for optical slicing, and without time-consuming 3D image reconstruction.
(Original title: Chinese Academy of Sciences developed a real-time laser microscope to obtain a patent for invention
ã€China Instrument Network Instrument Development】 The Super Resolution Imaging Team of the State Key Laboratory of Transient Optics and Photonics, Xi'an Institute of Optics and Precision Mechanics, Chinese Academy of Sciences, recently announced the successful development of a two-photon excitation laser scanning real-time stereo microscope for the first time based on binocular vision. The combination of stereomicroscopy and high resolution two-photon excitation laser scanning fluorescence microscopy enables high-speed stereoscopic imaging of three-dimensional fluorescence samples. At the same time, the research results also obtained the national invention patent authorization (Patent No. ZL201210384895.4).