Ultramicroscopic observation of recombinant adenoassociated virus type 2 on the surface of formvarcarbon coated copper grids under different relative humidity and incubation time using negative stain transmission electron microscopy

Abstract

The purpose of this investigation was to compare the effects of different relative humidity (RH) on the microcosmic conformation of the recombinant AAV-2 virion at 22°C. rAAV-2 virions prepared on copper grid were placed in a high, middle or low RH cabinet and incubated for 72, 48 and 24 h, respectively. The rAAV-2 virions were observed by transmission electron microscope and the values of major axis length, minor axis length and ellipticity of the rAAV-2 virions were obtained using the IMS cell image
analysis system. After incubation for 48 and 72 h, the major axis length and minor axis length of the rAAV-2 virion started to rapidly decrease in high RH. Conversely, the axis lengths rapidly increased in low RH. Then, the ellipticity of the rAAV-2 virion would almost tend to approach the identical value of
0.9 for 48 and 72 h incubations in high RH. The results suggest that the rAAV-2 virion tended to favor a smaller, round, more stable conformation in high RH compared to low RH which implied that the rAAV-2 virion was probably prone to living in high relative humidity conditions.