Comparative Observation of the Recombinant Adeno-Associated Virus 2 Using Transmission Electron Microscopy and Atomic Force Microscopy

Abstract

Adeno-associated virus (AAV) is a defective, nonpathogenic human parvovirus, which coinfects with a helper adenovirus or herpes virus. AAV's unique characteristics have made it an appealing vector system for gene delivery. AAV or recombinant AAV (rAAV) has been widely detected using negative stain transmission electron microscopy (TEM) but little has been detected using atomic force microscopy (AFM). In this article, we used AFM and TEM to observe the recombinant AAV-2 (rAAV-2) virus particles and applied statistical analysis to the AFM and TEM images. The results indicated that the rAAV-2 particle was a slightly elliptic particle close to round when it was detected by TEM (the mean length of major and minor axes of rAAV-2 particles was 24.77 ± 1.78 nm and 21.84 ± 1.57 nm, respectively), whereas when detected by AFM, the rAAV-2 particle was almost round. Even though the dimensions of the rAAV-2 particle exhibited a polymorphous distribution via off-line particle analysis of AFM, most of the rAAV-2 particles had a mean diameter of approximate 22.04 nm, which was similar to the results obtained by TEM. The results above suggested that AFM was important for accurately determining the average dimensions and distributions of virus particles.