Nanoscope 5 multimode 8 afm

TEM and AFM analysis was carried out at the Korea Basic Science Institute. For TEM analysis, conidia were fixed in 2.5% glutaraldehyde in 0.1 M phosphate, washed three times with 0.1 M phosphate, post-fixed in 1% osmium tetroxide, incubated for 1 h in 0.1 M phosphate, and dehydrated for 15 min in a graded methanol series from 50% to 100%. Samples were embedded in Epon resin 812. The sections were examined with a Tecnai G2 Spirit Twin Bio-Transmission Electron Microscope (FEI, Hillsboro, OR, USA), with an accelerating voltage of 120 KV. Conidial surfaces were analyzed by a Nanoscope V Multimode 8 AFM (Bruker, Santa Barbara, CA, USA). Conidia were immobilized by mechanically trapping them into porous polycarbonate membranes. After filtering a concentrated suspension of conidia, the filter was rinsed with deionized water, carefully cut, and attached to a metallic puck using double-sided sticky tape. Images were performed in soft tapping mode using a silicon AFM probe (k = 42 N/m, f = 320 kHz, NCHR, Nanoworld, Neuchâtel, Switzerland).