Multiphoton Microscopy

1. Front Matter

   Pages i-xii

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2. Building a Two-Photon Microscope Is Easy

   • Spencer LaVere Smith

   Pages 1-16

3. Using Multiphoton Imaging for Targeted Electrophysiological Recording and Live Cell Imaging of Fluorescently Labeled Neurons from Isolated Retinas

   • Qiang Chen, Wei Wei

   Pages 17-31

4. Two-Photon Neurotransmitter Uncaging for the Study of Dendritic Integration

   • Alexandra Tran-Van-Minh, Nelson Rebola, Andreas Hoehne, David A. DiGregorio

   Pages 33-64

5. Two-Photon Glutamate Uncaging to Study Structural and Functional Plasticity of Dendritic Spines

   • Ivar S. Stein, Travis C. Hill, Won Chan Oh, Laxmi K. Parajuli, Karen Zito

   Pages 65-85

6. Two-Photon Fluorescence Imaging of Visually Evoked Glutamate Release Using iGluSnFR in the Mouse Visual System

   • Bart G. Borghuis

   Pages 87-109

7. Imaging Neuronal Signal Transduction Using Multiphoton FRET-FLIM

   • Paul R. Evans, Long Yan, Ryohei Yasuda

   Pages 111-130

8. Combining Multiphoton Excitation Microscopy with Fast Microiontophoresis to Investigate Neuronal Signaling

   • Espen Hartveit, Margaret Lin Veruki

   Pages 131-159

9. Multiphoton Excitation Microscopy for the Reconstruction and Analysis of Single Neuron Morphology

   • Espen Hartveit, Bas-Jan Zandt, Margaret Lin Veruki

   Pages 161-194

10. Serial Multiphoton Tomography and Analysis of Volumetric Images of the Mouse Brain

    • Denise M. O. Ramirez, Apoorva D. Ajay, Mark P. Goldberg, Julian P. Meeks

    Pages 195-224

11. Studying a Light Sensor with Light: Multiphoton Imaging in the Retina

    • Thomas Euler, Katrin Franke, Tom Baden

    Pages 225-250

12. Multiphoton Ca2+ Imaging of Astrocytes with Genetically Encoded Indicators Delivered by a Viral Approach

    • Rune Enger, Rolf Sprengel, Erlend A. Nagelhus, Wannan Tang

    Pages 251-277

13. Two-Photon Imaging of Dendritic Calcium Dynamics In Vivo

    • Lucy M. Palmer

    Pages 279-295

14. Voltage Imaging with ANNINE Dyes and Two-Photon Microscopy

    • Christopher J. Roome, Bernd Kuhn

    Pages 297-334

15. High-Speed, Random-Access Multiphoton Microscopy for Monitoring Synaptic and Neuronal Activity in 3D in Behaving Animals

    • Mate Marosi, Gergely Szalay, Gergely Katona, Balázs Rózsa

    Pages 335-362

16. Back Matter

    Pages 363-364

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This volume covers the latest techniques and strategies used in multi-photon excitation (MPE) microscopy. Chapters in this book cover the fundamentals of MPE microscopy as applied to both in vitro and in vivo experimental systems; information on how to combine MPE microscopy with targeted electrophysiological recordings, calcium imaging, and transmembrane voltage imaging; methods to investigate cellular and large-scale neural morphology; signaling in astrocytes; and ways to use MPE microscopy to study the retina. In Neuromethods series style, chapters include the kind of detail and key advice from the specialists needed to get successful results in your laboratory.


Comprehensive and thorough, Multiphoton Microscopy is a valuable resource for both expert and novice researchers interested in expanding their knowledge and research in this rapidly developing field.

• calcium imaging
• STED microscopy
• multiphoton uncaging
• neuronal signal transduction
• membrane potential imaging
• Biological Microscopy

“I recommend this exciting book, replete with full-color images, because of its comprehensive, tested protocols for the study of live neural tissue at the cellular and the subcellular levels.” (Barry R. Masters, Optics & Photonics News, osa-opn.org, March 5, 2020)

• Department of Biomedicine, University of Bergen, Bergen, Norway

  Espen Hartveit

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