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Immunofluorescence
Published in Guy Cox, Fundamentals of Fluorescence Imaging, 2019
A polyclonal antiserum contains a mixture of different antibody isotypes as well as nonsense immunoglobulins that are normally present in the rabbit serum. Natural antibodies binding to the tissue section may occur in the antiserum as a result of unrecognized, prior antigenic stimulation. The strong staining of the epidermis, and moderate staining of vascular endothelial cells, fibroblasts, sweat glands, and arrector pili muscles of human skin described by Kobayashi et al. [6] after using some rabbit polyclonal antisera to lysozyme, myoglobin, S100 protein, and normal rabbit serum is probably due to the presence of anti-intermediate filament antibody, including an anti-keratin antibody. Such contaminating antibodies may co-purify with the specific antibody and cause false positive staining in immunofluorescence testing. Natural antibodies are usually found at low concentration and dilution may remove interference [7, 8]. It is generally accepted to use absorption controls where the antibody has been preabsorbed with the immunogen; however, this procedure does not eliminate the possibility that the antibody reacts with another protein in the tissue. Because antibodies recognize a relatively small component of an antigen, they can cross-react with similar epitopes on other antigens, but usually with less affinity [4, 9]. These low-affinity antibodies have weaker binding than specific antibodies. By incubating at low temperature, selection for the higher affinity specific antibody binding is preferred.
Early diagnosis of dental pathologies by front face fluorescence (FFF) and laser-induced breakdown spectroscopy (LIBS) with principal component analysis (PCA)
Published in Instrumentation Science & Technology, 2022
Rihem Nouir, Imen Cherni, Hassen Ghalila, Sami Hamzaoui
To identify correlations between concentrations of minerals, trace elements, and fluorescence peaks, Spearman’s rank correlation was applied to all measurements. Several tests were made involving the P4 and P5 peaks which correspond to vitamin C, S100 protein family grouping, vitamin D, and collagen. The choice of the P4 and P5 peaks is dictated by the numerous studies carried out to understand the relationship between the elemental concentrations with the vitamin concentrations. Our presentation was limited to the mineral and trace element correlations with the P5 peak because they provide the most significant results although the results with the P4 peak are similar.