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Imaging Local Ca2+ Signals in Cultured Mammalian Cells
09:30

Imaging Local Ca2+ Signals in Cultured Mammalian Cells

Published on: March 3, 2015

Ca2+-sensitive fluorescent dyes and intracellular Ca2+ imaging.

Martin D Bootman1, Katja Rietdorf, Tony Collins

  • 1Babraham Institute, Babraham, Cambridge, CB22 3AT, United Kingdom. martin.bootman@babraham.ac.uk

Cold Spring Harbor Protocols
|February 5, 2013
PubMed
Summary
This summary is machine-generated.

This guide explains how to use calcium imaging with fluorescent indicators to study cellular signals. It highlights potential challenges and considerations for researchers new to this technique.

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Imaging Local Ca2+ Signals in Cultured Mammalian Cells
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Applications of Spatio-temporal Mapping and Particle Analysis Techniques to Quantify Intracellular Ca2+ Signaling In Situ
09:34

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Single-Cell Calcium Imaging for Studying the Activation of Calcium Ion Channels
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Single-Cell Calcium Imaging for Studying the Activation of Calcium Ion Channels

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Area of Science:

  • Cellular Biology
  • Biophysics
  • Biochemistry

Background:

  • Calcium (Ca2+) signaling is crucial for numerous cellular processes.
  • Fluorescent indicators offer a noninvasive method for real-time Ca2+ measurement.
  • Targetable indicators enable domain-specific and multiplexed Ca2+ signal analysis.

Purpose of the Study:

  • To provide novice researchers with essential insights into using fluorescent Ca2+ indicators.
  • To outline key considerations and potential pitfalls in Ca2+ imaging experiments.
  • To facilitate the effective application of Ca2+ indicators in diverse research settings.

Main Methods:

  • Utilizing Ca2+-sensitive fluorescent indicators for cellular imaging.
  • Employing indicators with varying affinities, brightness, and spectral properties.
  • Targeting indicators to specific subcellular compartments for localized measurements.

Main Results:

  • Fluorescent Ca2+ indicators allow for the study of acute Ca2+ changes.
  • Indicator versatility supports exploration of spatiotemporal Ca2+ dynamics.
  • Multiplexing Ca2+ readouts with other cellular functions is achievable.

Conclusions:

  • Proper selection and application of fluorescent Ca2+ indicators are vital for accurate signal detection.
  • Understanding indicator characteristics and experimental context minimizes potential pitfalls.
  • This approach enhances the study of complex Ca2+ signaling pathways.