A method is described for converting video signals of analog scanning electron microscopes (SEMs) into digital images of high quality. A plug-in card commercially available for personal computers is used for the on-line analog/digital conversion. A Windows application program written by the authors, together with low-level software drivers supplied with the plug-in card, allow digital images to be recorded, to be displayed simultaneously on the computer monitor and to be saved as a file in a standardized format. Compared to conventional photographic images obtained from the SEM camera system, the digital images possess superior sharpness of outline, excellent image definition, diminished noise and well-defined grey-scale tones. This method provides SEM images of high quality for less than $1000 from most older analog SEMs. In addition, the advantages of digital image processing can be applied to analog SEMs, including contrast enhancement, digital filtering and multichannel recording.
A method for detecting glycoconjugates on cell surfaces in scanning electron microscopy is described. Terminal saccharides were specifically recognized by a lectin conjugated to biotin, and, after incubation with an anti-biotin antibody conjugated to colloidal gold, silver enhancement was used to produce deposits large enough to be detected in standard scanning electron microscopes. Secondary electron images revealed the ultrastructure of the tissue investigated, while backscattered electron images showed the distribution of lectin binding sites. Using digital recording and processing, the two channels were combined in colour-encoded images. The new method brings together lectin histochemistry and scanning electron microscopy and thus allows the three-dimensional distribution of glycoconjugates to be analysed at an ultrastructural level.
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