Abstract
The empirical demonstration of insulin by the basic (cationic) dye Victoria Blue 4R (V4R; Ivić 1969) shows the character of a histochemical reaction (Wohlrab et al. 1985), while the chemism of the reaction is not quite understood yet. Aim of this investigation: Will the spectral behaviour of V4R in visible light be influenced by oxidized or nonoxidized insulin?Dependent on the concentration, V4R shows in aqueous solution 2 absorption maxima (λ = 697 and 558 nm), which represents monomers and dimers of the dye. With increasing dye concentration (200 μmol/l), V4R forms dimers and even higher polymers. Constant V4R concentration (200 μmol/l) results with increasing concentration of oxidized insulin (4 to 16 μmo1/l) in a reduction of extinction, while the extinction at λ = 558 nm (dimers) is more decreased than at λ = 597 nm (monomers). Nonoxidized insulin has no remarkable influence on the absorption behaviour of V4R. Extinction measurements on V4R stained B-cells of islets of Langerhans after preoxidation of the section resulted in a main absorption maximum at λ = 554 nm.Conclusion: Depending on the concentration, the dye V4R is associated with the oxidized insulin in aqueous solution, which is also indicated by the occurrence of an isosbestic point in the curve behaviour. This is expressed by the establishment of a concentration-dependent equilibrium between the dye V4R and the oxidized insulin (change in the dissociation and aggregation behaviour respectively of the dye V4R). The determined main absorption maximum (λ = 554 nm) in the biological material points in the same way to interactions between the stain and oxidized nsulin.
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