Automation of 3D reconstruction of neural tissue from large volume of conventional serial section transmission electron micrographs.
We describe an approach for automation of the process of reconstruction of neural tissue from serial section transmission electron micrographs. Such reconstructions require 3D segmentation of individual neuronal processes (axons and dendrites) performed in densely packed neuropil. We first detect neuronal cell profiles in each image in a stack of serial micrographs with multi-scale ridge detector. Short breaks in detected boundaries are interpolated using anisotropic contour completion formulated in fuzzy-logic framework. Detected profiles from adjacent sections are linked together based on cues such as shape similarity and image texture. Thus obtained 3D segmentation is validated by human operators in computer-guided proofreading process. Our approach makes possible reconstructions of neural tissue at final rate of about 5 microm3/manh, as determined primarily by the speed of proofreading. To date we have applied this approach to reconstruct few blocks of neural tissue from different regions of rat brain totaling over 1000microm3, and used these to evaluate reconstruction speed, quality, error rates, and presence of ambiguous locations in neuropil ssTEM imaging data.
Janelia Farm Research Campus, Howard Hughes Medical Institute, Ashburn, VA 20147, USA. mishchenkoy@janelia.hhmi.org
Yuriy
Y
Mishchenko
D000818:Animals; D001331:Automation / Q000379:methods*; D001369:Axons / Q000648:ultrastructure; D021621:Imaging, Three-Dimensional / Q000379:methods*; D046529:Microscopy, Electron, Transmission / Q000295:instrumentation* / Q000379:methods*; D009474:Neurons / Q000648:ultrastructure*