RGB color coded images in scanning electron microscopy of biological surfaces

We present here a methodological approach for the creation of color images in scanning electron microscopy by processing grayscale images taken simultaneously from at least three different detectors in a scanning electron microscope. The final color images are then produced by merging together those grayscale images in RGB color space. We show the images from non-conductive standard sample together with those obtained from real microbiological samples. The first one represents a microbial biofilm naturally grown on fiber glass filter. The other shows individual Bacillus subtilis cells from batch culture. All the image handling was done in open source image processing software ImageJ or GNU Image Manipulation Program (Gimp) or, alternatively, in proprietary AnalySis 3.2 Pro software processing suite.