Division of Structural Biology, Wellcome Trust Centre for Human Genetics, University of Oxford, Roosevelt Drive, Oxford OX3 7BN, UK; Department of Biochemistry, University of Oxford, South Parks Road, Oxford OX1 3QU, UK.
Division of Structural Biology, Wellcome Trust Centre for Human Genetics, University of Oxford, Roosevelt Drive, Oxford OX3 7BN, UK.
Curr Opin Chem Biol. 2014 Jun;20(100):86-91. doi: 10.1016/j.cbpa.2014.05.007. Epub 2014 Jun 19.
Studying biological structures with fine details does not only require a microscope with high resolution, but also a sample preparation process that preserves the structures in a near-native state. Live-cell imaging is restricted mostly to the field of light microscopy. For studies requiring much higher resolution, fast freezing techniques (vitrification) are successfully used to immobilize the sample in a near-native state for imaging with electron and X-ray cryo-microscopy. Fluorescence cryo-microscopy combines imaging of vitrified samples with the advantages of fluorescence labeling of biological structures. Technical considerations as well as the behavior of fluorophores at low temperatures have to be taken into account for developing or adapting super-resolution methods under cryo conditions to exploit the full potential of this technique.
用精细细节研究生物结构不仅需要具有高分辨率的显微镜，还需要能够将结构在近乎自然状态下保存的样品制备过程。活细胞成像是光显微镜领域的主要方法。对于需要更高分辨率的研究，快速冷冻技术（玻璃化）成功地用于将样品固定在近乎自然的状态，以便使用电子和 X 射线冷冻显微镜进行成像。荧光冷冻显微镜将玻璃化样品的成像与生物结构的荧光标记的优势相结合。在开发或适应低温条件下的超分辨率方法以充分利用该技术的潜力时，必须考虑技术因素以及荧光团在低温下的行为。
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