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- W2405519281 abstract "Super-resolution microscopy systems have become more prominent in fluorescence imaging with many researchers interested in implementing these technologies. The three main types of super-resolution systems, structured illumination, serial localization and emission depletion, are expensive, complex and vary in their capabilities and platforms. The resolution gained in these different systems ranges from two- to ten-fold beyond that of the diffraction limit, and in practice is also dependent upon optimal sample preparation. Limited personal experience with these novel techniques can make it difficult to know which system will be the best purchase option for an institution. As one of the few facilities running both Stochastic Optical Reconstruction Microscopy (Nikon STORM) and 3-D Structured Illumination Microscopy (3D-SIM) (API OMX) systems, we are uniquely positioned to compare and benchmark these two different super-resolution techniques in practice using a well-characterized biological sample.The desmosome is a well-characterized biological structure with parallel cytoplasmic plaques on either side of the cell-cell junction. Each plaque component lines up at a distance from the plasma membrane that has been previously mapped by immunoelectron microscopy. Since resolution is defined as the ability to distinguish adjacent elements as separate components, the labeling of different plaque components is thus ideal for benchmarking the resolution achieved using our super-resolution systems. Two parallel fluorescent lines indicate resolution of the two protein domains, while a single line indicates that the resolution limit has been reached. For example, our results confirm predictions that the two bands of desmoplakin C-termini, ∼130 nm separation, can be resolved by both 3D-SIM and STORM, while the desmoplakin N-termini, only ∼50nm separation, can be resolved by STORM but not by 3D-SIM. Work is now in progress to maximize the resolution of each system by optimizing sample preparation and labeling, and to compare fixed and permeabilized cells with on-section staining." @default.
- W2405519281 created "2016-06-24" @default.
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- W2405519281 date "2013-05-01" @default.
- W2405519281 modified "2023-09-23" @default.
- W2405519281 title "Benchmarking the Resolution of 3D-SIM and STORM Using a Well-Characterized Biological System" @default.
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