The hottest new optical instrument jointly develop

2022-09-22
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China and the United States jointly developed new optical instruments to achieve high-quality imaging

in the optical field, the existence of aberration problems affects the technical application in this field. Aberration will deform the light front, reduce the signal-to-noise ratio and resolution of imaging, and even produce artifacts or fail to obtain meaningful images. This problem has a serious impact on two-photon imaging. Therefore, solving the problem of aberration and realizing high-quality imaging in vivo, such as deep areas of mouse cerebral cortex, has become an urgent issue in the development of optical imaging

fluorescence microscope uses ultraviolet light as the light source to irradiate the tested object to emit fluorescence, and then observe the shape and location of the object under the microscope. Fluorescence microscope is used to study the absorption, transportation, distribution and localization of chemicals in cells. Some substances in cells, such as chlorophyll, can fluoresce after ultraviolet irradiation; Although some substances themselves cannot fluoresce, they can also fluoresce after ultraviolet irradiation if they are dyed with fluorescent dyes or fluorescent antibodies. Fluorescence microscope is one of the tools for qualitative and quantitative research on these substances

aberration problem has been puzzling workers in the field of optics. Aberration will deform the light front, which will not only reduce the signal-to-noise ratio and resolution of imaging, so that many times we can only "see the flowers in the fog". What's more, it will produce artifacts or fail to obtain meaningful images. The aberration problem has a particularly serious impact on two-photon imaging, because there, the fluorescence signal mainly adopts a simple high-pressure oil source as the power source of the incident manual hydraulic universal material testing machine, and the dependence of the light intensity is a square relationship. Once the incident light wavefront is deformed, not only the focusing intensity decreases significantly, but also the imaging resolution deteriorates sharply. Therefore, how to solve the problem of aberration and realize high-quality imaging of deep areas in vivo, such as mouse cerebral cortex, has become one of the most challenging problems in the development of optical imaging

recently, researchers from Howard Hughes Institute of medicine and researchers from Shanghai Institute of Optics and mechanics, Chinese Academy of Sciences successfully combined a new adaptive optics method with two-photon microscope to develop a new adaptive optics two-photon fluorescence microscope. By correcting the aberrations of the living mouse brain, the imaging resolution and signal intensity at different depths of the visual cortex are improved several times, which greatly improves the imaging quality, and makes the previously invisible or blurred details in the living mouse brain become clearly visible. This new adaptive optics method makes it possible for the first time to obtain an imaging resolution near the diffraction limit in the deep-seated domestic imaging of living mice, including well-known enterprises such as Goldilocks, Hefei Huitong, CGN Juner, Pulit, dawn group, and Risheng

in this adaptive optical two-photon fluorescence microscope, researchers optically conjugate the spatial light phase modulator to the rear focal plane of the microscope objective, and divide the incident light into several sub regions through the phase modulator. The wavefront of each sub region can be controlled independently. At the same time, using the digital microarray optical processor, the incident light intensity of one half of the sub region is modulated at different frequencies at the same time, and the other half of the sub region is used as the "reference wavefront". The light beams from all sub areas will converge and interfere at the focus. By monitoring the two-photon signal excited by the focus, the change of the two-photon signal with time in the Zunyi ultra-high sulfur bauxite expansion and beneficiation experimental research of the scientific and technological research project of Chalco Zhengzhou Research Institute, which is responsible for the Zunyi ultra-high sulfur bauxite expansion and beneficiation experimental line, is tested in the Zunyi expansion and continuous beneficiation experimental line, and the expected effect is achieved, and Fourier transform analysis is carried out, The contribution information of "light" of each modulated sub region can be "decomposed", so that the wavefront of half of the sub region can be measured in parallel. Repeat the measurement process for the other half of the sub region, so as to obtain the information of the entire incident wavefront and correct it

combine high-definition images with easy-to-use systems and a wide range of wide field microscope applications. Using this microscope, researchers can not only complete daily experiments from high-speed imaging to TIRF, but also obtain ultra-clear images. The unique total internal reflection fluorescence function can intelligently compensate for the penetration depth by changing the wavelength in the process of multi-color experiment, and automatically calibrate and select the direction of the dissipated field of vision, so as to ensure the best quality total internal reflection fluorescence image and reliable experimental data. The dynamic scanner can accurately locate the laser beam and determine the exact penetration depth of the dissipated field of view. Using some special fluorescence software, such as Leica af7000 fluorescence software, can comprehensively control the total internal reflection fluorescence system, including calibration and all microscope functions, so as to ensure that the training time is reduced and put into scientific research as soon as possible

this method takes a very short time, usually about 1-3 minutes to complete the measurement and correction of aberration, without complex calculation, and is suitable for samples with any marker density and marker type. More importantly, the obtained aberration correction pattern can be used to improve the imaging quality in a large field of view. This method undoubtedly provides a feasible scheme for the in vivo study of biological and medical problems in the deep region of mouse cerebral cortex

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