ONE Microscopy: Imaging Proteins at the Nanoscale
ONE microscopy is a new imaging technology developed by researchers from the University Medical Center Göttingen, able to expand specimens tenfold and allow the visible inspection of individual protein shapes with 1-nm resolution using conventional light microscopes.
An unprecedented subnanometer resolution for imaging single proteins or small molecular complexes has now been achieved with a facile nanoscale expansion microscopy, one-step version. One step further from achieving a more ideal concept of fluorescence microscopy than ever before is possible: pushing the dramatic nanoscale boundary. Researchers can now see the intricate shapes of membrane and soluble proteins at unprecedented detail, up to 1-nanometer resolution, by expanding specimens tenfold and using conventional fluorophores with standard light microscopes. Not only does this new approach bridge the gap between structural biology and microscopy, but it also has tremendous promise for analyzing dynamic protein behavior and more intricate biological systems.
Capturing Dynamic Protein Conformational Changes.
ONE microscopy utilizes a groundbreaking process that avoids the resolution limit of light microscopy. By physically expanding the sample, researchers can obtain information about the fine structures that were unattainable with a traditional imaging method. The method involves tagging proteins with conventional fluorophores and recording video sequences that enable an examination of fluorescence fluctuations. It allows the straightforward visualization of real-time conformational changes.
One of the more significant accomplishments would be the imaging of calmodulin, a protein which has large structural changes upon binding with Ca2+. The visuals seen via this method provide a higher resolution, proving this technique has tremendous scope in understanding the dynamics of proteins at molecular level.
Biological origin samples are linked to gel matrices through Acryloyl-X, where X10 gel forms and is then homogenized through proteinase K digestion or autoclaving in alkaline buffers. Through repeated washing in distilled water, samples are mounted in a specific chamber, where fluorophores are separated spatially, independent fluctuation detection is possible through repeated images, up to 3,000 times, through different imaging systems such as confocal and epifluorescence, among others. These fluctuations are then processed using the SRRF algorithm on the ONE platform to provide super-resolved images that can be overlaid with cryo-EM data for enhanced visualisation.
Clinical Applications: New Horizons in Diagnosing Diseases
Beyond basic research, ONE microscopy promises much in the way of translational applications. The technology was used to characterise protein aggregation in samples of cerebrospinal fluids withdrawn from patients suffering from Parkinson’s disease. This has the potential to be a groundbreaking approach in the diagnosis and monitoring of diseases that are characterized by protein misfolding and aggregation with all their attendant potential for providing clear insight into disease progression, perhaps even leading to earlier diagnoses.
Advantages and Future Prospects
ONE microscopy closes not only the resolution gap between light microscopy and higher-resolution structural biology techniques, such as cryo-electron microscopy but also provides a more accessible, versatile platform for researchers. The use of conventional microscopes makes the technique more adaptable and cost-effective, opening its adoption to even wider research labs and clinical environments.
This technique’s power in capturing fine details of individual shapes of proteins and observing dynamic changes in real-time makes it the discovery tool scientists need the most. Its potential is huge – not just in furthering frontiers of understanding molecular biology but also in changing the whole landscape of disease diagnosis, particularly neurodegenerative diseases. Further refining it would provide a cornerstone for research, in addition to its potential use in clinical applications and unlock a deeper understanding of complex biological systems.
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