High-Resolution Calcium Imaging Method for Local Calcium Signaling

Methods Mol Biol. 2019;1929:27-39. doi: 10.1007/978-1-4939-9030-6_3.

Abstract

All eukaryotic cells respond to extracellular signals in a physiologically meaningful way. For multicellular organisms, physiologically relevant cooperation is only possible, if cell-cell communication works properly. Here, the extracellular signals must be translated into intracellular signals that ultimately result in cellular responses. This process is termed signal transduction or signaling. Ca2+ signaling has been developed in almost all eukaryotic cells. The cellular components used for this highly versatile signaling system are often termed "Ca2+ toolbox". Besides Ca2+ pumps and Ca2+-binding proteins, the Ca2+ channels that are located in the plasma membrane and intracellular membranes and the Ca2+-mobilizing second messengers are major players in shaping the four-dimensional nature of Ca2+ signals.Here, we report on methodological developments to acquire and analyze cellular Ca2+ signals with high temporal and spatial resolution with specific focus on (1) photobleaching of Ca2+ indicators at high acquisition rate, (2) determination of system noise and spatiotemporal detection limits, and (3) image processing.

Keywords: Ca2+ microdomain cell signaling; Ca2+ signaling; Deconvolution; High-resolution imaging; Live cell imaging.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Calcium / analysis*
  • Calcium Channels / metabolism
  • Calcium Signaling*
  • Calcium-Binding Proteins / metabolism
  • Cell Communication
  • Cell Membrane / metabolism*
  • Mice
  • Microscopy, Fluorescence
  • Photobleaching
  • Second Messenger Systems
  • T-Lymphocytes / metabolism*

Substances

  • Calcium Channels
  • Calcium-Binding Proteins
  • Calcium