Method and device for correlated micro-imaging of frozen light microscope and frozen electron microscope

Abstract

The invention relates to a method and a device for correlated micro-imaging of a frozen light microscope and a frozen electron microscope, in particular to a method and a device for correlated imaging of a frozen light microscope and a frozen transmission electron microscope, including an optical microscope frozen object stage device used for optics-electron correlated micro-imaging and a corresponding correlated imaging method. By adopting the method and the device, optical micro-imaging at extra-low temperature (below minus 170 degrees) can be achieved, and frozen electron micro-imaging of the same samples in the same areas can also be achieved. The method and the device can be widely used to correlate various types of inverted optical microscopes and most transmission electron microscopes at the market, can achieve correlated imaging fast, conveniently and accurately, prevent ice crystals from forming on surfaces of the samples to the maximum extent, avoid problems of grid deformation and carbon film fracture, and can be widely applied to related research of cytobiology, neurobiology, biological medicines, etc.

Description

technical field [0001] The present invention relates to equipment and methods for microscopic imaging, in particular to the associated imaging of optical microscopy and electron microscopy, especially the associated imaging of cryo-optic microscopy and cryo-transmission electron microscopy, including optical-electron microscopy associated imaging. Microscope cryo-stage setup and corresponding associated imaging methods. Background technique [0002] Cryo-photocorrelation microscopy imaging: a method that can perform optical microscopic detection of samples labeled with optical probes at extremely low temperatures (below -170 ° C, capable of maintaining the fine structure information of samples in close to physiological states), and A technique that enables high-resolution electron microscopic imaging detection of the same area of ​​the sample. Cryo-photonic correlation microscopy combines the advantages of modern fluorescence microscopy and cryo-electron microscopy to make ...

AI Technical Summary

Problems solved by technology

The problems of this system are: during the cryo-light microscope imaging process, the sample is exposed to the air, which causes the sample to be easily polluted by ice crystals; the cartridge is small, and only depends on its own gravity to fit the cryo-associated platform, so it is less stable under cryo-conditions Poor, causing the sample to shake a lot during the imaging process; it is only suitable for the Polara electron microscope of FEI Company, and the scope of promotion and use is small

In addition, the cryo-correlation platform developed by the research group of Wolfgang Baumeister of the Max Planck Institute in Germany and the research group of Abraham J.Koster of Leiden University in the Netherlands uses a self-made sample holder to carry the electron microscope sample carrier grid. After the cryo-light microscope imaging is completed, It is necessary to remove the sample grid from the sample holder, transfer it to the sample holder or sample holder of the corresponding electron microscope, and then transfer it to the cryo-electron microscope for imaging, which leads to the need for the sample grid to be transferred multiple times: on the one hand, multiple times The transfer increases the difficulty of correlation; on the other hand, the repeated exposure of frozen samples to air and boiling liquid nitrogen makes the samples unusable due to ice crystal contamination, grid deformation and carbon film breakage

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