Volume 99, Issue 12, Pages (December 2010)

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Volume 99, Issue 12, Pages 3923-3929 (December 2010) Analysis of Cav1.2 and Ryanodine Receptor Clusters in Rat Ventricular Myocytes David R.L. Scriven, Parisa Asghari, Meredith N. Schulson, Edwin D.W. Moore Biophysical Journal Volume 99, Issue 12, Pages 3923-3929 (December 2010) DOI: 10.1016/j.bpj.2010.11.008 Copyright © 2010 Biophysical Society Terms and Conditions

Figure 1 (A) Stereo pair, 6° separation, of a ventricular cell labeled for RyR (red) and Cav1.2 (green); colocalized voxels are in white. Scale bars are 5 μm in each dimension. (B) Indicated Z disks isolated and rotated 90° about the X axis; 12 planes deep in Y. (C) Magnified images of the clusters highlighted in B are displayed one plane at a time through the width of the clusters (i–vi). Biophysical Journal 2010 99, 3923-3929DOI: (10.1016/j.bpj.2010.11.008) Copyright © 2010 Biophysical Society Terms and Conditions

Figure 2 Single image planes taken from three cells labeled for RyR and Cav1.2. (A) Wide-field microscope (RyR, red; Cav1.2, green). Couplons traversing an entire sarcomere (arrow) and part of a sarcomere (double arrow) are indicated. (B and C) Confocal microscope, cells viewed end-on. In B, RyR is red and Cav1.2 is green; the colors are reversed in C. Colocalized voxels are always white. Three display paradigms are used: (i) voxel colocalization, (ii) colocalized voxels are removed from the image, and (iii) cluster colocalization. Single arrows point to rings of RyR, which are magnified in the insets; double arrows point to extradyadic clusters of RyR. Scale bar: 5 μm. Biophysical Journal 2010 99, 3923-3929DOI: (10.1016/j.bpj.2010.11.008) Copyright © 2010 Biophysical Society Terms and Conditions

Figure 3 Histograms showing the distribution of the NNDs between the indicated clusters of molecules. Data are from the 25 cells imaged on the confocal microscope. (A) RyR. (B) Cav1.2. (C) Couplons. (D) Extradyadic RyR and Cav1.2. (E) Extradyadic RyR. (F) Extradyadic Cav1.2. (G) Extradyadic RyR to couplon. (H) Extradyadic Cav1.2 to couplon. A total of 42,259 clusters of RyR and 26,587 clusters of Cav1.2 were analyzed. Bin size: 50 nm. Biophysical Journal 2010 99, 3923-3929DOI: (10.1016/j.bpj.2010.11.008) Copyright © 2010 Biophysical Society Terms and Conditions