Application of the Sterecon System for the Determination of Area, Volume, and Linear Distance of Cell Surface Topography of Secondary and Backscatter Electron Stereoscopic Images in Low-Voltage Field Emission SEM

Abstract

Extraction of quantitative information on surface topography in SEM can.be accomplished with, stereoscopic visual parallaxes. This method has received little attention in the last decade and to our knowledge has not been applied to high resolution secondary or backscatter electron images from field emission SEM. The STERECON system was developed for 3-D reconstruction of cells from stereo images obtained by high voltage EM. The present study describes the application of STERECON to quantitative determination of volume, surface area, and linear distance of topographical features in high resolution SEM.Trophozoites of the free-living protozoan, Hexamita inflata, and two bacterial species (Proteus mirabilus and Enterococcus faecalis) were fixed in 2.5-3.0% glutaraldehyde, dehydrated in ethanol, critical point dried in CO2, and coated with about 1 nm of platinum before examination at low kV (<4.0) in a Hitachi S-900 field emission SEM equipped with secondary (Se) and backscatter (YAG scintillator) detectors and photographed on Polaroid type 52 film using a stereo tilt of 8°.