Figure 7.

Simulation of accuracy of synapse detection by AT. (A) Simulated AT images using a previously segmented EM image data set [from Mishchenko et al. (2010)]. (A1) an example EM image; (A2) same image segmented into pre- and post-synaptic structures; (A3) segmented image was blurred to produce the same resolution as in the light microscopy of AT with 90% of structures removed to mimic the sparseness of fluorescent labeling of the AT images; (A4) The pixel size of A3 was adjusted to produce the same image as in the light microscopy in AT. Red indicates presynaptic structures, green postsynaptic, blue synaptophysin and red arrowheads indicates a synapse. Scale bar = 1 μm (B) Close-up of a simulated synapse showing postsynaptic structure (green) and simulated synaptophysin staining (blue). Note that the predicted synaptophysin staining exhibits an increasing intensity gradient toward the synaptic contact as found experimentally (Figure 10; Scale bar = 0.5 μm). (C) False positive rate in simulations of AT using different light imaging isotropic spatial resolutions. Black dashed line indicates resolution of the light imaging in the current study. Blue dotted line is an extrapolation of simulated data to infinite resolution.