3d reconstruction of vzv infected cell nuclei and pml nuclear cages by serial section array scanning electron microscopy and electron tomography3 d重建带状疱疹感染细胞核和pml核笼子连续切片阵列扫描电子显微镜和电子断层扫描.pdfVIP

3D Reconstruction of VZV Infected Cell Nuclei and PML Nuclear Cages by Serial Section Array Scanning Electron Microscopy and Electron Tomography 1 2 2 3 2 1 Mike Reichelt *, Lydia Joubert , John Perrino , Ai Leen Koh , Ibanri Phanwar , Ann M. Arvin 1 Departments of Pediatrics and Microbiology Immunology, Stanford University School of Medicine, Stanford, California, United States of America, 2 Cell Sciences Imaging Facility, Stanford University School of Medicine, Stanford, California, United States of America, 3 Stanford Nanocharacterization Laboratory, Geballe Laboratory for Advanced Materials, Stanford University, Stanford, California, United States of America Abstract Varicella-zoster virus (VZV) is a human alphaherpesvirus that causes varicella (chickenpox) and herpes zoster (shingles). Like all herpesviruses, the VZV DNA genome is replicated in the nucleus and packaged into nucleocapsids that must egress across the nuclear membrane for incorporation into virus particles in the cytoplasm. Our recent work showed that VZV nucleocapsids are sequestered in nuclear cages formed from promyelocytic leukemia protein (PML) in vitro and in human dorsal root ganglia and skin xenografts in vivo. We sought a method to determine the three-dimensional (3D) distribution of nucleocapsids in the nuclei of herpesvirus-infected cells as well as the 3D shape, volume and ultrastructure of these unique PML subnuclear domains. Here we report the development of a novel 3D imaging and reconstruction strategy that we term Serial Section Array-Scanning Electron Microscopy (SSA-SEM) and its application to the analysis of VZV-infected cells and these nuclear PML cages. We show that SSA-SEM permits large volume imaging an