Slides were mounted with Histomount (pH 6.5 7.1, TAAB laboratories) and viewed using a Zeiss Axioskop microscope equipped with x20/0.5.
Ultrathin sections of 70 90 nm were cut with an ultramicrotome, EM UC7 (Leica), stained with uranyl acetate plus lead citrate, and viewed using a JEM1010 transmission electron microscope (JEOL, Japan).
Whole leaves from 5 6-week-old plants expressing the GRX-roGFP2 redox sensor38, inoculated with 107 cfu/mL avirulent PstAvrB or 10 mM MgCl2 as a control, were cut at the level of the petiole, gently placed on a slide and covered with a coverslip, and viewed using a Nikon Ti-E inverted fluorescence microscope (Nikon Corp., Tokyo, Japan) with a CFI Plan Fluor 4 × N.A. 0.13 dry objective.
Images were captured and viewed using a Q imaging Retiga-SRV camera and QCapture Pro 6.0 software (Q Imaging, Surrey, British Columbia, Canada).
Transgenic strains of C. elegans were mounted on 2% agarose pads and viewed using a Leica confocal microscope fitted with Nomarski optics.
Scanning EM samples were sputter coated with a Gold-Palladium alloy, sectioned and viewed using a Hitachi Model S-3000N Scanning EM.
The cells were again washed three times with PBS, counter-stained with DAPI, and viewed using a 63× objective on a Zeiss Axiovert 200 M microscope.
The sections were then washed, stained with Hoechst 33258 (1 µg/ml) for 2 min, and viewed using a confocal microscope (FluoView™ FV300, OLYMPUS).
Transmission EM samples were heavy metal stained with Uranyl Acetate and Lead Citrate, sectioned and viewed using a Hitachi Model H-7600 Transmission EM.
After washing, the coverslips were mounted using Fluorsave (Calbiochem) and viewed using a Zeiss Axiovert 135 microscope as previously described [1].
The slides were mounted in Fluoromount-G (Southern Biotechnology) containing counter staining dye 4',6-diamidino-2-phenylindole (Sigma) and viewed using a Zeiss Axiophot epifluorescent microscope.
