FIGURE 2.1. In vivo two-photon imaging of individual neurons and neuronal populations.


In vivo two-photon imaging of individual neurons and neuronal populations. (A) Schematic of a two-photon microscope setup for in vivo imaging in the brain. Beam control includes beam expansion and adjustment of laser intensity. Fluorescence can be detected in multiple color channels using photomultiplier tubes (PMTs). Image stacks are acquired using a software-controlled focusing device, for example, a piezoelectric focusing element. (B) The typical view during the experiment is the top view (xy view; top row). To obtain side views (xz or yz views) of neurons, maximum-intensity side projections are computed from stacks of hundreds of images (bottom row). The neuron on the left was filled with a fluorescent calcium indicator through a whole-cell patch pipette. The focal plane of the xy image was just above the main bifurcation of the apical dendrite. The neuronal population on the right was visualized in negative contrast by bulk injection of the bright red fluorescent dye Alexa-594 into the extracellular space. Note that pyramidal cells can be identified by their apical dendritic trunks. Scale bars 20 μm.

From: Chapter 2, Two-Photon Functional Imaging of Neuronal Activity

Cover of In Vivo Optical Imaging of Brain Function
In Vivo Optical Imaging of Brain Function. 2nd edition.
Frostig RD, editor.
Boca Raton (FL): CRC Press/Taylor & Francis; 2009.
Copyright © 2009, Taylor & Francis Group, LLC.

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