Photon counting, censor corrections, and lifetime imaging for improved detection in two-photon microscopy.
نویسندگان
چکیده
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.
منابع مشابه
Innovative Methodology Photon counting, censor corrections, and lifetime imaging for improved detection in two-photon microscopy
Jonathan D. Driscoll, Andy Y. Shih, Satish Iyengar, Jeffrey J. Field, G. Allen White, Jeffrey A. Squier, Gert Cauwenberghs, and David Kleinfeld Department of Physics, University of California at San Diego, La Jolla, California; Department of Statistics, University of Pittsburgh, Pittsburgh, Pennsylvania; Department of Physics, Colorado School of Mines, Golden, Colorado; and Department of Bioeng...
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متن کاملPhoton counting , censor corrections , and lifetime imaging for 1 improved detection in two - photon microscopy 2 3
3 Jonathan D. Driscoll, Andy Y. Shih, Satish Iyengar, Jeffrey J. Field, 4 G. Allen White, Jeffrey A. Squier, Gert Cauwenberghs and David Kleinfeld 5 6 Department of Physics, University of California at San Diego, La Jolla, CA 7 Department of Statistics, University of Pittsburgh, Pittsburgh, PA 8 Department of Physics, Colorado School of Mines, Golden, CO 9 Department of Bioengineering, Universi...
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For each experimental run, we assume that the number of photocurrent events, i.e., detected fluorescent emission events, N , is a Poisson random variable with mean α. If N = n ≥ 1, then let W1, . . . ,Wn be the times of the N emissions; we assume that each Wi is exponentially distributed and that the emission times are independent of the number of emissions. For all calculations below, our cloc...
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Time-correlated single photon counting (TCSPC) is a sensitive, robust and precise technique to measure luminescence decays.[1] For fluorescence microscopy, it is often implemented with beam scanning and single point detectors, but we have implemented a camera-based wide-field TCSPC method. We employ a photon counting image intensifier in combination with a 1 MHz frame rate CMOS camera, thus com...
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ورودعنوان ژورنال:
- Journal of neurophysiology
دوره 105 6 شماره
صفحات -
تاریخ انتشار 2011