• Svoboda Karel
• Yasuda Ryohei

Neuron

• Publish Date: Jun 2006
• ISSN: 0896-6273
• Volume: 50
• Issue: 6
• Pages: 823-39
• Medium: Print
• Language: English
• Citation (JAMA): Svoboda Karel, Yasuda Ryohei, et al. Principles of Two-photon Excitation Microscopy and Its Applications to Neuroscience.. Neuron Jun 2006;50:823-39

Abstract

The brain is complex and dynamic. The spatial scales of interest to the neurobiologist range from individual synapses (approximately 1 microm) to neural circuits (centimeters); the timescales range from the flickering of channels (less than a millisecond) to long-term memory (years). Remarkably, fluorescence microscopy has the potential to revolutionize research on all of these spatial and temporal scales. Two-photon excitation (2PE) laser scanning microscopy allows high-resolution and high-sensitivity fluorescence microscopy in intact neural tissue, which is hostile to traditional forms of microscopy. Over the last 10 years, applications of 2PE, including microscopy and photostimulation, have contributed to our understanding of a broad array of neurobiological phenomena, including the dynamics of single channels in individual synapses and the functional organization of cortical maps. Here we review the principles of 2PE microscopy, highlight recent applications, discuss its limitations, and point to areas for future research and development.

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