Abstract
Using a very stable (Δx & Δy < 0.7 nm RMS) dSTORM imaging system we have endeavored to accurately determine the position of the Type 2 Ryanodine (RyR2) on the surface of the cardiomyocyte. After labeling the tetramers with an Alexa 647 fluorophore, we collected millions of blinks from each sample to properly define the receptor positions. We then used Delaunay triangulation followed by an edge cull to identify clusters of RyR2 whose blinks were no more than a given distance (usually 50 nm) from each other. Combining this approach with a measure of blink density, we were able to isolate the signal from the tetramers and, in some cases, visualize both their position and orientation. The distribution of the tetramers in a cluster is irregular, but not random, and does not follow any distribution. Grouping of two or more tetramers is common. These results are in agreement with previously published tomographic data. Estimates of the number of tetramers per cluster are made by combining super-resolution and tomographic data.
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