Three-dimensional (3D) reconstruction is the last and an essential step toward high-resolution structural determination in single-particle cryo-electron microscopy (cryoEM). We have implemented a new algorithm for reconstructing 3D structures of macromolecular complexes with icosahedral symmetry from cryoEM images. Icosahedral symmetry-adapted functions (ISAFs) are used to interpolate structural factors in the reciprocal space to generate a 3D reconstruction in spherical coordinates. In our implementation, we introduced a recursive method for deriving higher order ISAFs from three lower order seed functions. We demonstrate improvements of our new method in both the noise suppression and the effective resolution in 3D reconstruction over the commonly used Fourier-Bessel synthesis method introduced by Crowther et al. three decades ago. Our 3D reconstruction method can be extended to macromolecular complexes with other symmetry types and is thus likely to impact future high-resolution cryoEM single-particle reconstruction efforts in general. (c) 2007 Elsevier Inc. All rights reserved.