Immunofluorescence microscopy studies of the granular and keratin layers of normal human skin

Granular and keratin layer fluorescence of normal human skin has been found to be very different, depending on whether the indirect or the direct fluorescent antibody technique is used. These layers fluoresce more brightly with the indirect than with the direct technique. These differences were interpreted as being caused by in vitro binding of antibodies such as anti-stratum corneum antibodies. The purpose of this study was to see if these differences were, in part, artifacts. on serum dilution, the appearance with the indirect technique consistently converted to that of the direct technique, but no microscopic structural differences were found between the skins subjected to the different technical procedures. Therefore, it was concluded that the indirect appearance is caused only by antibodies binding to the keratin and granular layers and is not a methodological artifact.