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Evaluation of Two Fluorescent Dyes used in Immunofluorescent Microscopy for the Detection of Proteinaceous Binding Media in Wall Paintings

Abstract

The following study sought to compare the efficacy of Qdot antibody conjugates to FITC antibody conjugates for secondary staining in the identification of proteinaceous binding media in wall paintings using immunofluorescence microscopy (IFM) and to further explore limitations of the technique as a result of pigment binder interactions. A protocol was developed for the identification of egg-based media in wall paintings using FITC for IFM, but no protocol was developed for the Qdot antibodies. Three pigments were used to explore pigment binder interactions and their effects on the results of IFM (vermilion, hematite, and lead white). Hematite combined with egg tempera was found to have reduced signal strength when compared to vermilion in the same binder. Lead white was found to be problematic due to enhanced autofluorescence of the organic binding media, making it difficult to differentiate positive and negative results.

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