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J Neurophysiol. 2011 Jun;105(6):3106-13. doi: 10.1152/jn.00649.2010. Epub 2011 Apr 6.

Photon counting, censor corrections, and lifetime imaging for improved detection in two-photon microscopy.

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1
Department of Physics, University of California at San Diego, La Jolla, CA 92093-0374, USA.

Abstract

We present a high-speed photon counter for use with two-photon microscopy. Counting pulses of photocurrent, as opposed to analog integration, maximizes the signal-to-noise ratio so long as the uncertainty in the count does not exceed the gain-noise of the photodetector. Our system extends this improvement through an estimate of the count that corrects for the censored period after detection of an emission event. The same system can be rapidly reconfigured in software for fluorescence lifetime imaging, which we illustrate by distinguishing between two spectrally similar fluorophores in an in vivo model of microstroke.

PMID:
21471395
PMCID:
PMC3118755
DOI:
10.1152/jn.00649.2010
[Indexed for MEDLINE]
Free PMC Article
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