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Cellular functions are dictated by the intricate motion of proteins in membranes that span across a scale of nanometers to micrometers, within a time-frame of microseconds to minutes. However, this rich parameter or space is inaccessible using fluorescence microscopy, although it is within reach of interferometric scattering (iSCAT) particle tracking. The new iSCAT technique is, however, highly sensitive to single and unlabelled proteins, thereby causing non-specific background staining as a substantial challenge during cellular imaging.
In a recent study, Richard W. Taylor and colleagues at the interdisciplinary departments of Physics and Biology in Germany developed a new image processing approach to overcome this difficulty. They used the method to track the transmembrane epidermal growth factor receptor (EGFR) with nanometer scale precision in three dimensions (3-D). The technique allowed imaging across microseconds to minutes. The scientists provided examples of nanoscale motion and confinement using the method to image ubiquitous processes such as diffusion in plasma membranes, transport in filopodia and rotational motion during endocytosis. The results are now published in Nature Photonics.
Progress - Fluorescence - Microscopy - Scientists - Events
While steady progress in fluorescence microscopy has allowed scientists to monitor cellular events at the nanometer scale, a great deal still remains to be accomplished with advanced imaging systems. The challenges of fluorescence microscopy occurred due to the finite emission rate of a fluorescent source (dye molecule or semiconductor quantum dot), where too few photon emissions during a very small time-frame prevented effective or prolonged imaging. The central difficulty of scattering-based microscopy...
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