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J Neurophysiol (May 9, 2007). doi:10.1152/jn.01169.2006
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Submitted on November 1, 2006
Accepted on May 8, 2007

Methods for voltage-sensitive dye imaging of rat cortical activity with high signal-to-noise ratio

Michael Thomas Lippert1, Kentaroh Takagaki1, Weifeng Xu1, Xiaoying Huang1, and Jian-young Wu2*

1 Department of Physiology and Biophysics, Georgetown University, Washington, District of Columbia, United States
2 Physiology & Biophysics, Georgetown University, Washington, District of Columbia, United States

* To whom correspondence should be addressed. E-mail: wuj{at}georgetown.edu.

We describe methods to achieve high sensitivity in voltage-sensitive dye (VSD) imaging from rat barrel and visual cortices in vivo with the use of a blue dye RH1691 and a high dynamic range imaging device (photodiode array). With an improved staining protocol and an off-line procedure to remove pulsation artifact, the sensitivity of VSD recording is comparable to that of local field potential recording from the same location. With this sensitivity, one can record from ~500 individual detectors, each covering an area of cortical tissue 160 µm in diameter (total imaging field ~4 mm in diameter) and a temporal resolution of 1,600 frames/s, without multiple-trial averaging. We can record 80 to 100 trials of intermittent 10 s trials from each imaging field before the VSD signal reduces to one half of its initial amplitude due to bleaching and wash-out. Taken together, the methods described in this report provide a useful tool for visualizing evoked and spontaneous waves from rodent cortex.




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