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This volume looks at the latest developments in studying calcium signaling. The chapters in this book discuss topics such as in vitro and in vivo analyses in model systems; methods to measure Ca2+ in different subcellular compartments; single Ca2+ channels; methods to measure cellular ROS and ATP; and the functionality of the ATP synthase. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

Comprehensive and cutting-edge, Calcium Signalling: Methods and Protocols is a valuable resource that covers both conceptual and methodological viewpoints to aid beginners and experts in furthering their studies in the developing field of calcium homeostasis research.

Calcium Signalling: Methods and Protocols

Preface...
Table of Contents...
Contributing Authors...

1.High-Throughput Screening using Photoluminescence Probe to Measure Intracellular Calcium Levels
Simona Feno, Giulia di Marco, Agnese di Mario, Halenya Monticelli, and Denis Vecellio Reane

2.Exploiting Cameleon Probes to Investigate Organelles Ca2+ Handling
Luisa Galla, Paola Pizzo, and Elisa Greotti 

3.Measuring Ca2+ Levels in Subcellular Compartments with Genetically-Encoded GFP-Based Indicators
Mattia Vicario and Tito Cali 

4.Methods to Measure Intracellular Ca2+ Concentration using Ca2+-Sensitive Dyes
Sofia Zanin, Elisa Lidron, and Giorgia Pallafacchina 

5.MCU Regulation in Lipid Bilayer and Electrophysiological Recording
Vanessa Checchetto and Ildiko Szabo

6.Electrophysiological Characterization of Calcium-Permeable Channels using Planar Lipid Bilayer
Vanessa Checchetto and Ildiko Szabo

7.Patch Clamp Analysis of the Mitochondrial Calcium Uniporter
Vivek Garg and Yuriy Y. Kirichok

8.In Vivo Light Sheet Fluorescence Microscopy of Calcium Oscillations in Arabidopsis thaliana
Neli Romano Armada, Fabrizio Gandolfo Doccula, Alessia Candeo, Gianluca Valentini, Alex Costa, and Andrea Bassi

9.Ex Vivo Measurements of Ca2+ Transients in Intracellular Compartments of Skeletal Muscle Fibres by Means of Genetically Encoded Probes
Gaia Gherardi and Cristina Mammucari

10.Imaging Intracellular Ca2+ in Cardiomyocytes with Genetically Encoded Fluorescent Probes
Antonio Campo and Marco Mongillo

11.In Vivo Monitoring of Ca2+ Uptake into Subcellular Compartments of Mouse Skeletal Muscle
Rüdiger Rudolf, Sofie Trajanovska, David Grant Allen, and Tullio Pozzan

12.TRPML1/TFEB-Dependent Regulation of Lysosomal Exocytosis 
Simone Di Paola and Diego L. Medina

13.Ca2+-Dependent Regulation of TFEB and Lysosomal Function
Simone Di Paola and Diego L. Medina

14.Employing Transgenic Parasite Strains to Study the Ca2+ Dynamics in the Human Malaria Parasite Plasmodium falciparum
Lucas Borges-Pereira and Célia R. S. Garcia

15.Calcium Imaging of Stored Operated Calcium (Ca2+) Entry (SOCE) in HEK293 Cells using Fura-2
Martin Johnson 

16.Optogenetic Interneuron Stimulation and Calcium Imaging in Astrocytes
Gabriele Losi, Anna Maria Lia, Marta Gomez-Gonzalo, Micaela Zonta, and Giorgio Carmignoto

17.Measuring Calcium and ROS by Genetically Encoded Protein Sensors and Fluorescent Dyes
Christine Gibhardt, Adina Vultur, and Ivan Bogeski

18.Assessing Calcium-Stimulated Mitochondrial Bioenergetics using the Seahorse XF96 Analyzer
Jennifer Wettmarshausen and Fabiana Perocchi

19.Determination of ATP, ADP, AMP Levels by Reversed-Phase High Performance Liquid Chromatography in Cultured Cells
Michela Menegollo, Isabella Tessari, Luigi Bubacco, and Gyorgy Szabadkai

20.Purification of Functional F-ATP Synthase from Blue Native PAGE
Chiara Galber, Giulia Valente, Sophia von Stockum, and Valentina Giorgio