Classification of heterogeneous electron microscopic projections into homogeneous subsets

The co-existence of different states of a macromolecular complex in samples used by three-dimensional electron microscopy (3D-EM) constitutes a serious challenge. The single particle method applied directly to such heterogeneous sets is unable to provide useful information about the encountered conformational diversity and produces reconstructions with severely reduced resolution. One approach to solving this problem is to partition heterogeneous projection set into homogeneous components and apply existing reconstruction techniques to each of them. Due to the nature of the projection images and the high noise level present in them, this classification task is difficult. A method is presented to achieve the desired classification by using a novel image similarity measure and solving the corresponding optimization problem. Unlike the majority of competing approaches, the presented method employs unsupervised classification (it does not require any prior knowledge about the objects being classified) and does not involve a 3D reconstruction procedure. We demonstrate a fast implementation of this method, capable of classifying projection sets that originate from 3D-EM. The methodʹs performance is evaluated on synthetically generated data sets produced by projecting 3D objects that resemble biological structures.