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J Neurophysiol (March 14, 2007). doi:10.1152/jn.01178.2006
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01178.2006v1
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Submitted on November 6, 2006
Accepted on March 10, 2007

Combined voltage and calcium epifluorescence imaging in vitro and in vivo reveals subthreshold and suprathreshold dynamics of mouse barrel cortex

Thomas Berger1, Aren Jacobus Borgdorff2, Sylvain Crochet2, Florian Neubauer1, Sandrine Lefort2, Bruno Fauvet2, Isabelle Ferezou2, Alan Carleton3, Hans-R. Rudolf Luscher1, and Carl CH Petersen2*

1 University of Bern, Institute of Physiology, Bern, Switzerland
2 Laboratory of Sensory Processing, Brain and Mind Institute, Lausanne, Switzerland
3 Flavor Perception Group, Brain and Mind Institute, Lausanne, Switzerland

* To whom correspondence should be addressed. E-mail: carl.petersen{at}epfl.ch.

Cortical dynamics can be imaged at high spatiotemporal resolution with voltage-sensitive dyes and calcium-sensitive dyes. We combined these two imaging techniques using epifluorescence optics together with whole-cell recordings to measure the spatiotemporal dynamics of activity in the mouse somatosensory barrel cortex in vitro and in the supragranular layers in vivo. The two optical signals reported distinct aspects of cortical function. Voltage-sensitive dye fluorescence varied linearly with membrane potential and was dominated by subthreshold postsynaptic potentials, whereas the calcium-sensitive dye signal predominantly reflected local action potential firing. Combining voltage-sensitive dyes and calcium-sensitive dyes allowed us to monitor the synaptic drive and the spiking activity of a given area at the same time in the same preparation. The spatial extent of the two dye signals was different, with voltage-sensitive dye signals spreading further than calcium-sensitive dye signals, reflecting broad subthreshold and narrow suprathreshold receptive fields. Importantly, the signals from the dyes were differentially affected by pharmacological manipulations, stimulation strength and depth of anesthesia. Combined voltage-sensitive dye and calcium-sensitive dye measurements can therefore be used to specify the temporal and spatial relationships between subthreshold and suprathreshold activity of the neocortex.




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