Real-Time Investigation of Plasma Membrane Deformation and Fusion Pore Expansion Using Polarized Total Internal Reflection Fluorescence Microscopy

• DOI: 10.1007/978-1-4939-0944-5_18
• Publication Date: 2014-06-19T14:54:21Z
• Authors: Daniel R. Passmore; Tejeshwar Rao; Arun Anantharam
• Keywords: Microscopy, Fluorescence; Cell Membrane; Animals; Microscopy, Polarization; Membrane Fusion; Cells, Cultured; Endocytosis; Exocytosis

Polarized Total Internal Reflection Fluorescence Microscopy (pTIRFM) allows for real-time observation of plasma membrane deformations. The technique provides insights into the dynamics of biological processes requiring rapid and localized changes in membrane shape. Such processes include exocytosis, endocytosis, cytokinesis, and cell motility. In this chapter, we describe how to implement a polarization-based TIRF imaging system to monitor exocytosis in adrenal chromaffin cells.