Figure 4.
Figure 4. Alignment and idealization of junction images. (A) EM freeze-fracture image of a surface junction with vertical shading. (B) Illustration of the alignment stage in quantitative analysis. A grid representing the underlying array of RYR channels is superimposed to the image in A. Yellow squares represent V channels, and red squares represent C channels. The cyan polygon traces the putative borders of the couplon. (C) The outcome of idealization of the image in A. Orange circles represent CaVs interacting with V channels, cyan circles are CaVs interacting with C channels, and purple circles are particles that cannot be placed in interactions with RYR channels. (D and E) Additional examples of successful analysis. C and D are obtained from junctions in BC3H1 cells. E is from an image of a frog surface junction. The spatial scale is provided by the tetrads, which are ∼30 nm on each side.

Alignment and idealization of junction images. (A) EM freeze-fracture image of a surface junction with vertical shading. (B) Illustration of the alignment stage in quantitative analysis. A grid representing the underlying array of RYR channels is superimposed to the image in A. Yellow squares represent V channels, and red squares represent C channels. The cyan polygon traces the putative borders of the couplon. (C) The outcome of idealization of the image in A. Orange circles represent CaVs interacting with V channels, cyan circles are CaVs interacting with C channels, and purple circles are particles that cannot be placed in interactions with RYR channels. (D and E) Additional examples of successful analysis. C and D are obtained from junctions in BC3H1 cells. E is from an image of a frog surface junction. The spatial scale is provided by the tetrads, which are ∼30 nm on each side.

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