New visualization technology is found to be eliminated by electron microscope
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In situ imaging techniques are challenging because there are many limitations, but now they can be overcome: (i) The low temperature focusing example beam provides insight into frozen cells and tissues that are too thick to be viewed directly by transmission electron microscopy. (ii) Direct observation improves the image quality of the frozen transmission electron microscope. (iii) The recently developed voltage phase plate (VPP) improves the phase of especially low spatial frequencies, and there is no defocusing to compare, so that the obtained image can directly display the structural condition.
Hela cells labeled β-tubulin by green fluorescent protein and mCherry labeled histone 2B. The analysis of three-dimensional images near the nucleus by cryo-electron tomography shows that macromolecular complexes in the nucleus and nuclear membrane can be directly visualized.
In order to provide better quantitative detection, the study examined 80S ribosomes, which produced images with an average resolution of 28 Ã…, and then the endoplasmic reticulum-bound ribose according to their spatial arrangement in the endoplasmic reticulum. The body (per 143 capsules) produced an image with a resolution of 35Ã…. The average and classification of ribosomes by sub-X-ray faults reflects the natural structure and organization of this cytoplasmic translation machine.
Macromolecular complexes are generally not strictly determined because their structure is altered by function. The large dynamic structural analysis of the nuclear pore complexes in this study shows that this new technique can detect differences in individual complexes.
Cryo-ET can be used to visualize in situ visualization of previously difficult to honed structures such as nucleosome chains and core layered filaments.