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  • Title: Automatic identification of crossovers in cryo-EM images of murine amyloid protein A fibrils with machine learning.
    Author: Weber M, Bäuerle A, Schmidt M, Neumann M, Fändrich M, Ropinski T, Schmidt V.
    Journal: J Microsc; 2020 Jan; 277(1):12-22. PubMed ID: 31859366.
    Abstract:
    Detecting crossovers in cryo-electron microscopy images of protein fibrils is an important step towards determining the morphological composition of a sample. Currently, the crossover locations are picked by hand, which introduces errors and is a time-consuming procedure. With the rise of deep learning in computer vision tasks, the automation of such problems has become more and more applicable. However, because of insufficient quality of raw data and missing labels, neural networks alone cannot be applied successfully to target the given problem. Thus, we propose an approach combining conventional computer vision techniques and deep learning to automatically detect fibril crossovers in two-dimensional cryo-electron microscopy image data and apply it to murine amyloid protein A fibrils, where we first use direct image processing methods to simplify the image data such that a convolutional neural network can be applied to the remaining segmentation problem. LAY DESCRIPTION: The ability of protein to form fibrillary structures underlies important cellular functions but can also give rise to disease, such as in a group of disorders, termed amyloid diseases. These diseases are characterised by the formation of abnormal protein filaments, so-called amyloid fibrils, that deposit inside the tissue. Many amyloid fibrils are helically twisted, which leads to periodic variations in the apparent width of the fibril, when observing amyloid fibrils using microscopy techniques like cryogenic electron microscopy (cryo-EM). Due to the two-dimensional projection, parts of the fibril orthogonal to the projection plane appear narrower than parts parallel to the plane. The parts of small width are called crossovers. The distance between two adjacent crossovers is an important characteristic for the analysis of amyloid fibrils, because it is informative about the fibril morphology and because it can be determined from raw data by eye. A given protein can typically form different fibril morphologies. The morphology can vary depending on the chemical and physical conditions of fibril formation, but even when fibrils are formed under identical solution conditions, different morphologies may be present in a sample. As the crossovers allow to define fibril morphologies in a heterogeneous sample, detecting crossovers is an important first step in the sample analysis. In the present paper, we introduce a method for the automated detection of fibril crossovers in cryo-EM image data. The data consists of greyscale images, each showing an unknown number of potentially overlapping fibrils. In a first step, techniques from image analysis and pattern detection are employed to detect single fibrils in the raw data. Then, a convolutional neural network is used to find the locations of crossovers on each single fibril. As these predictions may contain errors, further postprocessing steps assess the quality and may slightly alter or reject the predicted crossovers.
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