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

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 impro...

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Published inJournal of neurophysiology Vol. 105; no. 6; pp. 3106 - 3113
Main Authors Driscoll, Jonathan D, Shih, Andy Y, Iyengar, Satish, Field, Jeffrey J, White, G Allen, Squier, Jeffrey A, Cauwenberghs, Gert, Kleinfeld, David
Format Journal Article
LanguageEnglish
Published United States American Physiological Society 01.06.2011
Subjects
Analog-Digital Conversion
Animals
Brain - cytology
Cell Death
Diagnostic Imaging - methods
Green Fluorescent Proteins - genetics
Innovative Methodology
Interneurons - physiology
Luminescent Measurements - methods
Mice
Mice, Inbred C57BL
Mice, Transgenic
Microtubule-Associated Proteins - metabolism
Photons
Signal Processing, Computer-Assisted
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Summary: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.
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ISSN:0022-3077
1522-1598
DOI:10.1152/jn.00649.2010