The purpose of this study was to identify pitfalls in the Gram staining technic that limit its diagnostic value. In our clinical experience, gram-positive organisms were often decolorized too easily. Factors have been identified that alter the susceptibility of gram-positive organisms to decolorization in the Gram staining technic. The age of the bacterial culture, the preparation of the smear, the fixation technic, and the mordant have an important influence on the ease with which gram-positive organisms are decolorized. On the basis of these studies, a more reliable and reproducible Gram staining technic has been developed for the diagnosis of surgical infections.
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