caa a22 4500 469030895 CHVBK 20180323132743.0 cr unu---uuuuu 170328e19920501xx s 000 0 eng 10.1007/BF01046838 doi (NATIONALLICENCE)springer-10.1007/BF01046838 Confocal microscopical analysis of epithelial cell heterogeneity in mouse Peyer's patches [Elektronische Daten] [P. James, C. Rossetti, M. Smith, D. Cremaschi] Summary: Cyanine dye fluorescence and alkaline phosphatase activities have been compared directly by confocal microscopy in a wide variety of cells present in the follice-associated epithelium of the mouse Peyer's patch to test the hypothesis that antigen-transporting M cells have a low membrane potential. In order to make these comparisons it was first necessary to equilibrate living tissue with the membrane potential sensitive dye DIOC5(3), fix with glutaraldehyde and then incubate the fixed tissue with naphthol AS-BI phosphate, a substrate which is hydrolysed by alkaline phosphatase present in the luminal membrane of these epithelial cells. Naphthol AS-BI produced by this reaction, is then coupled to Fast Red TR diazonium salt at the site of hydrolysis. Selecting the 488 nm wavelength of the argon laser source then allows one to measure alkaline phosphatase activities as Fast Red absorbance and membrane potentials by DIOC5(3) fluorescence. Results obtained show a linear correlation between membrane potential and alkaline phosphatase activity. Relative lack of alkaline phosphatase activity, determined in fixed tissue, has been used previously to identify antigen-transporting M cells (Smithet al., 1987). The present work shows that it is now possible to recognize these cells in living tissue by measurement of DIOC5(3) fluorescence. The possible importance of this finding in providing a way to study cell surface-antigen interactions taking place in living tissue is discussed. Chapman & Hall, 1992 James P. AFRC Institute of Animal Physiology and Genetics Research, Babraham, CB2 4AT, Cambridge, UK aut Rossetti C. Dipartimento di Fisiologia e Biochimica Generali, Universita degli Studi di Milano, Via Celoria 26, 20133, Milano, Italy aut Smith M. AFRC Institute of Animal Physiology and Genetics Research, Babraham, CB2 4AT, Cambridge, UK aut Cremaschi D. Dipartimento di Fisiologia e Biochimica Generali, Universita degli Studi di Milano, Via Celoria 26, 20133, Milano, Italy aut The Histochemical Journal Kluwer Academic Publishers 24/5(1992-05-01), 243-250 0018-2214 24:5<243 1992 24 10735 https://doi.org/10.1007/BF01046838 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/BF01046838 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- James P. AFRC Institute of Animal Physiology and Genetics Research, Babraham, CB2 4AT, Cambridge, UK aut NATIONALLICENCE 700 1- Rossetti C. Dipartimento di Fisiologia e Biochimica Generali, Universita degli Studi di Milano, Via Celoria 26, 20133, Milano, Italy aut NATIONALLICENCE 700 1- Smith M. AFRC Institute of Animal Physiology and Genetics Research, Babraham, CB2 4AT, Cambridge, UK aut NATIONALLICENCE 700 1- Cremaschi D. Dipartimento di Fisiologia e Biochimica Generali, Universita degli Studi di Milano, Via Celoria 26, 20133, Milano, Italy aut NATIONALLICENCE 773 0- The Histochemical Journal Kluwer Academic Publishers 24/5(1992-05-01), 243-250 0018-2214 24:5<243 1992 24 10735 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer