Histochemical staining and estimation of zinc content in pancreatic β-cells by using of Dithizon and 8-para(toluenesulphonylamino)quinolin

Abstract

It is known that zinc in pancreatic β-cells take part in processes of formation of deposited form of insulin. Assessment of its contents allows to estimate functional state of β-cells. It is necessary to use a precise and high specific histochemical methods for identification of zinc in β-cells. Authors presented results of experimental improving of methods for qualitative and quantitative estimation of zinc content in pancreas tissue. Authors showed that: 1) decrease of thickness of sections of pancreas tissue 1,5 times — from 6–7 mcm to 4 mcm have best results for investigate of histotopography of zinc in islets using of Dithizon method; 2) similar result is obtained by decrease of dose of Dithizon njected to animals for 2–2,5 times; 3) quantitative estimation of zinc content is possible by using of histofluorimetric complex; 4) formation of a complex of dithizon with zinc in the form of the painted granules significantly improve results of analysis of histotopography of zinc in islet; 5) regarding preparing of frozen sections of pancreas tissue: best results were obtained using of temperature of freezing of tissue from -18o to – 22 oC; 6) more precise result were obtained using of additional method staining of zinc by TSH which is more sensitive for revealing of minimal concentration of zinc as 107-10-8. Thus, obtained results allow to ex- pand and significantly improve possibilities of histochemical Dithizon and TSH methods for estimate concentration and disposition of zinc in pancreatic islets.