Biological membranes are one of the most basic structures and regions of interest in cell biology. In the study of membranes, segment extraction is a well-known and difficult problem because of impeding noise, directional and thickness variability, etc. Recent advances in electron microscopy membrane segmentation are able to cope with such difficulties by training convolutional neural networks. However, because of the massive amount of features that have to be extracted while propagating forward, the practical usability diminishes, even with state-of-the-art GPU's. A significant part of these network features typically contains redundancy through correlation and sparsity. In this work, we propose a pruning method for convolutional neural networks that ensures the training loss increase is minimized. We show that the pruned networks, after retraining, are more efficient in terms of time and memory, without significantly affecting the network accuracy. This way, we manage to obtain real-time membrane segmentation performance, for our specific electron microscopy setup.
We present a fast algorithm for training MaxPooling Convolutional Networks to segment images. This type of network yields record-breaking performance in a variety of tasks, but is normally trained on a computationally expensive patch-by-patch basis. Our new method processes each training image in a single pass, which is vastly more efficient. We validate the approach in different scenarios and report a 1500-fold speed-up. In an application to automated steel defect detection and segmentation, we obtain excellent performance with short training times. To build the connectomics map of the brain, we developed a new algorithm that can automatically refine the Membrane Detection Probability Maps (MDPM) generated to perform automatic segmentation of electron microscopy (EM) images. To achieve this, we executed supervised training of a convolutional neural network to recover the removed center pixel label of patches sampled from a MDPM. MDPM can be generated from other machine learning based algorithms recognizing whether a pixel in an image corresponds to the cell membrane. By iteratively applying this network over MDPM for multiple rounds, we were able to significantly improve membrane segmentation results.  In the field of neuroanatomy, automatic segmentation of electron microscopy images is becoming one of the main limiting factors in getting new insights into the functional structure of the brain. We propose a novel framework for the segmentation of thin elongated structures like membranes in a neuroanatomy setting. The probability output of a random forest classifier is used in a regular cost function, which enforces gap completion via perceptual grouping constraints. The global solution is efficiently found by graph cut optimization. We demonstrate substantial qualitative and quantitative improvement over state-of the art segmentations on two considerably different stacks of ssTEM images as well as in segmentations of streets in satellite imagery. We demonstrate that the superior performance of our method yields fully automatic 3D reconstructions of dendrites from ssTEM data.  The analysis of microcircuitry (the connectivity at the level of individual neuronal processes and synapses), which is indispensable for our understanding of brain function, is based on serial transmission electron microscopy (TEM) or one of its modern variants. Due to technical limitations, most previous studies that used serial TEM recorded relatively small stacks of individual neurons. As a result, our knowledge of microcircuitry in any nervous system is very limited. We applied the software package TrakEM2 to reconstruct neuronal microcircuitry from TEM sections of a small brain, the early larval brain of Drosophila melanogaster. TrakEM2 enables us to embed the analysis of the TEM image volumes at the microcircuit level into a light microscopically derived neuro-anatomical framework, by registering confocal stacks containing sparsely labeled neural structures with the TEM image volume. We imaged two sets of serial TEM sections of the Drosophila first instar larval brain neuropile and one ventral nerve cord segment, and here report our first results pertaining to Drosophila brain microcircuitry. Terminal neurites fall into a small number of generic classes termed globular, varicose, axiform, and dendritiform. Globular and varicose neurites have large diameter segments that carry almost exclusively presynaptic sites. Dendritiform neurites are thin, highly branched processes that are almost exclusively postsynaptic. Due to the high branching density of dendritiform fibers and the fact that synapses are polyadic, neurites are highly interconnected even within small neuropile volumes. We describe the network motifs most frequently encountered in the Drosophila neuropile. Our study introduces an approach towards a comprehensive anatomical reconstruction of neuronal microcircuitry and delivers microcircuitry comparisons between vertebrate and insect neuropile.