A teflon culture dish for high-magnification microscopy and measurements in single cells
- 92 Downloads
A simple tissue culture dish is described which permits the use of oil immersion optics (x100) while maintaining access to cells in culture. In combination with a micro-CO2-incubator (Ince et al. 1983) long-term as well as short-term experiments can be performed under microscopical control. The teflon dish is re-usable, resistant to sterilization procedures, and easy to assemble. Standard glass coverslips with adherent cells are secured to the dish by an aluminium ring with a bayonet fitting. Both phasecontrast and interference-contrast microscopy with high magnification can be used. The dish is of particular use in electrophysiological investigations where the pathch-clamp technique is used and frequent exchange of cultures is required. With this unit, single ion channel activity in cultured human monocytes was measured. The dish has also been used in studies in which membrane potential measurements were performed with glass microelectrodes in small cells under culture conditions. The applications described include video time-lapse sequences of phagocytosis with microorganisms presented to mouse peritoneal macrophages with broken-tipped pipettes. This easy to use, multi-purpose dish offers the cell physiologist a valuable aid for the manipulation and observation of single cells in culture.
Key wordsTeflon culture dish Human monocytes and mouse peritoneal macrophages Patch clamp Video time-lapse microscopy
Unable to display preview. Download preview PDF.
- Hamill OP, Marty A, Neher E, Sakmann B, Sigworth FJ (1981) Improved patch-clamp techniques for high-resolution current recording from cells and cell-free membrane patches. Pflügers Arch 391:85–100Google Scholar
- Ince C, Ypey DL (1984) Membrane hyperpolarizations and ionic channels in cultured human monocytes. In: van Furth R (ed) Proceedings of the IVth International congress on mononuclear phagocytes. Martinus Nijhoff Publ., The Hague, The Netherlands (in press)Google Scholar
- Ince C, Ypey DL, Diesselhoff-Den Dulk MMC, Visser JAM, de Vos A, van Furth R (1983a) Micro-CO2-incubator for use on a microscope. J Immun Meth 60:269–275Google Scholar
- Ince C, Ypey DL, van Furth R, Verveen AA (1983b) Estimation of the membrane potential of cultured macrophages from the fast potential transient upon microelectrode entry. J Cell Biol 96:796–801Google Scholar
- Ince C, Leijh PCJ, Meijer J, van Bavel E, Ypey DL (1984) Oscillatory hyperpolarizations and resting membrane potentials of mouse fibroblast and mouse macrophage cell lines. J Physiol (Lond) 352:625–635Google Scholar
- Leijh PCJ, van den Barselaar MTh, van Zwet TL, Daha MR, van Furth R (1979) Requirement of extracellular complement for intracellular killing of microorganisms by human monocytes. J Clin Invest 63:772–784Google Scholar
- Matteson DR, Deutsch C (1984) K channels in T lymphocytes: a patch clamp study using monoclonal antibody adhesion. Nature 307:468–471Google Scholar
- Nijweide PJ, van Yperen-van Gent AS, Kawilarang-De Haas EWM, van der Plas A, Wassenaar AM (1982) Bone formation and calcification by isolated osteoblast-like cells. J Cell Biol 93:318–323Google Scholar
- Van Dijk AM, Wieringa PA, van der Meer M, Laird JD (1984) Mechanics of resting isolated single vascular smooth muscle cells from bovine coronary artery. Am J Physiol 246:C277-C287Google Scholar
- Van Furth R, Cohn ZA (1968) The origin and kinetics of mononuclear phagocytes. J Exp Med 128:415–435Google Scholar