Three-dimensional fluorescence microscopic imaging method based on optical tomography reconstruction strategy
A three-dimensional fluorescence and optical tomography technology, which is used in the analysis of fluorescence emission, diagnosis and diagnosis using tomography, etc. Imaging, enhance the effect of imaging depth
Pending Publication Date: 2022-01-21
FUDAN UNIV
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Therefore, the existing ultra-high-resolution fluorescence microscopy imaging technology is difficult to perform rapid and three-dimensional imaging at the whole-cell level, which cannot meet the growing needs of biomedical research.
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[0017] In order to make the technical means, creative features, goals and effects of the present invention easy to understand, a three-dimensional fluorescence microscopic imaging method based on the optical tomographic reconstruction strategy of the present invention will be described in detail below in conjunction with the embodiments and accompanying drawings.
[0018]
[0019] figure 1 It is a flowchart of a three-dimensional fluorescence microscopy imaging method based on an optical tomography reconstruction strategy in an embodiment of the present invention.
[0020] Such as figure 1 As shown, a three-dimensional fluorescence microscopy imaging method based on an optical tomography reconstruction strategy includes the following steps:
[0021] Step S1, using the photoswitchable fluorescent probe to perform fluorescence microscopic imaging on the cells of the biological tissue in the reconstruction area, and measure the fluorescence value ζ(r). With the intervention o...
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The invention provides a three-dimensional fluorescence microscopic imaging method based on an optical tomography reconstruction strategy, which is characterized by comprising the following steps of: S1, performing fluorescence microscopic imaging on cells of biological tissues in a reconstruction area by utilizing a light switchable fluorescence probe, and measuring a fluorescence value zeta(r); S2, obtaining a fluorescence value zeta(r) according to first-order Born approximation; S3, solving a Green function according to a diffusion equation; S4, in combination with boundary conditions, performing solving by utilizing a finite element method and a sparse reconstruction method to obtain three-dimensional distribution of the fluorescent probe in the cells at a certain imaging moment; and S5, generating a three-dimensional fluorescence localization microscopic image according to the calculated three-dimensional distribution superposition of the fluorescent probe in the cells at all imaging moments. According to the invention, an optical tomography reconstruction method and a super-resolution fluorescence microscopy technology are combined; rapid three-dimensional imaging is carried out by constructing a super-resolution fluorescence microscopy three-dimensional reconstruction model; and the spatial resolution of three-dimensional imaging is improved.
Description
technical field [0001] The invention relates to a three-dimensional fluorescence microscopic imaging method based on an optical tomography reconstruction strategy. Background technique [0002] As a new fluorescence microscopy imaging technology developed in recent years, Super-Resolution Fluorescence Microscopy (for example: Stochastic Optical Reconstruction Microscopy, STORM) and Photoactivated Localization Microscopy PALM (Photoactivated Localization Microscopy)] breaks through the optical diffraction limit and can increase the lateral spatial resolution to 20-50nm, that is, it can analyze the intracellular organelles of living cells at the single-molecule level (nanoscale). Or molecular structure imaging, which will help to understand the molecular mechanism of biological processes and the molecular basis of disease at the molecular level. [0003] With the development of biomedicine, researchers hope to observe the three-dimensional structure and spatial relationship o...
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IPC IPC(8): A61B5/00G06T17/20
CPCA61B5/0071A61B5/0073A61B5/0084A61B5/6848G06T17/205
Inventor 刘欣刘成成李博艺他得安
Owner FUDAN UNIV