A sub field of medical physics/molecular biology, thh overall aim of the effort is to improve the understanding of the molecular architecture of unknown or understudies biological systems, for example of the human sperm (flagellum component) using cryo-electron tomography and advanced image processing workflows.
In this example, men fertilization issues from accidents, excessive therapeutic radiation and pathological development in puberty call for scientists to solve the structures of key flagellar macromolecular complexes to understand the molecular mechanisms controlling sperm function both in health and disease.
Do you know about the current degree of effectiveness compared to other approaches and the reliability in bi8logical systems structure elucidation by cryo-electron tomography ?
Cryo-electron tomography (cryo-ET) is a technique that allows the study of the 3D structure of cells and tissues at near-native conditions⁴. It can provide unprecedented insights into the inner working of molecular machines in their native environment, as well as their functional relevant conformations and spatial distribution within biological cells or tissues¹.
The field of structure elucidation by cryo-ET has evolved rapidly in recent years, thanks to the advances in instrumentation, sample preparation, image processing, and data analysis. Some of the current trends and challenges in the field are:
- Achieving higher resolution and better contrast for cryo-ET images, by using phase plates, direct electron detectors, and improved alignment and reconstruction algorithms¹³.
- Developing new methods and protocols for cryo-ET sample preparation, such as cryo-focused ion beam milling, cryo-correlative light and electron microscopy, and cryo-electron microscopy of vitreous sections¹⁴.
- Applying cryo-ET to study a wide range of biological systems and processes, such as viruses, bacteria, organelles, cytoskeleton, membrane proteins, and macromolecular complexes¹²³.
- Integrating cryo-ET with other complementary techniques, such as mass spectrometry, X-ray crystallography, nuclear magnetic resonance, and computational modeling, to obtain a comprehensive understanding of the structure and function of biological macromolecules¹⁵.
- Disseminating and standardizing cryo-ET data and structures in public databases, such as the Protein Data Bank (PDB) and the Electron Microscopy Data Bank (EMDB), to facilitate data sharing and reproducibility⁵.
(1) Cryo-electron Tomography Sample Preparation - Thermo Fisher Scientific. https://www.thermofisher.com/us/en/home/electron-microscopy/life-sciences/sample-preparation/techniques/cryo-tomography.html.
(2) High-resolution in situ structure determination by cryo-electron .... https://www.nature.com/articles/s41596-021-00648-5.pdf.
(3) Cryo-electron tomography: 3-dimensional imaging of soft matter. https://pubs.rsc.org/en/content/articlehtml/2011/sm/c0sm00441c.
(4) JoF | Free Full-Text | Preliminary Structural Elucidation of β-(1,3 .... https://www.mdpi.com/2309-608X/7/2/120/htm.
(5) Evolution of standardization and dissemination of cryo-EM structures .... https://www.jbc.org/article/S0021-9258%2821%2900338-0/pdf.
(6) undefined. https://doi.org/10.1038/s41596-021-00648-5.
(7) undefined. https://doi.org/10.3390/jof7020120.
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