Three dimensional pixel super-resolution microscopic imaging method
A microscopic imaging and super-resolution technology, used in material excitation analysis, fluorescence/phosphorescence, etc., which can solve the problems of large field of view and high resolution that are difficult to be compatible.
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[0031] The three-dimensional structure of lung cells cultured in vitro is imaged by using the three-dimensional pixel super-resolution microscopic imaging method and slice light microscopic imaging technology of the present invention. Imaging settings were 4x acquisition objective + 10 µm thick illumination light sheet. According to this setting, in the obtained low-resolution three-dimensional image Bi, in the x-y direction, the size of a single pixel is the camera pixel size 6.45 μm / 4 = 1.61 μm, and in the z direction, the size of a single pixel is half the thickness of the light sheet, that is, 5 Micron. According to the Enquist sampling principle, the images correspond to raw lateral and axial resolutions of ~3.2 μm and ~10 μm, respectively. In the image acquisition, in order to achieve the super-resolution ratio of Ex=5, Ey=5, Ez=10, when generating 250 sets of low-resolution images, we selected an overscanning step size of 10000nm / 250=40nm. Next, the original image seq...
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