  We introduce a new rotationally invariant viewing angle classification method
for identifying, among a large number of Cryo-EM projection images, similar
views without prior knowledge of the molecule. Our rotationally invariant
features are based on the bispectrum. Each image is denoised and compressed
using steerable principal component analysis (PCA) such that rotating an image
is equivalent to phase shifting the expansion coefficients. Thus we are able to
extend the theory of bispectrum of 1D periodic signals to 2D images. The
randomized PCA algorithm is then used to efficiently reduce the dimensionality
of the bispectrum coefficients, enabling fast computation of the similarity
between any pair of images. The nearest neighbors provide an initial
classification of similar viewing angles. In this way, rotational alignment is
only performed for images with their nearest neighbors. The initial nearest
neighbor classification and alignment are further improved by a new
classification method called vector diffusion maps. Our pipeline for viewing
angle classification and alignment is experimentally shown to be faster and
more accurate than reference-free alignment with rotationally invariant K-means
clustering, MSA/MRA 2D classification, and their modern approximations.
