Real-time dynamic range and signal to noise enhancement in beam-scanning microscopy by
integration of sensor characteristics, data acquisition hardware, and statistical methods

David J. Kissick; Ryan D. Muir; Shane Z. Sullivan; Robert A. Oglesbee; Garth J Simpson

doi:10.1117/12.2008607

26 February 2013 Real-time dynamic range and signal to noise enhancement in beam-scanning microscopy by integration of sensor characteristics, data acquisition hardware, and statistical methods

David J. Kissick, Ryan D. Muir, Shane Z. Sullivan, Robert A. Oglesbee, Garth J Simpson

Author Affiliations +

Proceedings Volume 8657, Computational Imaging XI; 86570E (2013) https://doi.org/10.1117/12.2008607
Event: IS&T/SPIE Electronic Imaging, 2013, Burlingame, California, United States

Abstract

Despite the ubiquitous use of multi-photon and confocal microscopy measurements in biology, the core techniques typically suffer from fundamental compromises between signal to noise (S/N) and linear dynamic range (LDR). In this study, direct synchronous digitization of voltage transients coupled with statistical analysis is shown to allow S/N approaching the theoretical maximum throughout an LDR spanning more than 8 decades, limited only by the dark counts of the detector on the low end and by the intrinsic nonlinearities of the photomultiplier tube (PMT) detector on the high end. Synchronous digitization of each voltage transient represents a fundamental departure from established methods in confocal/multi-photon imaging, which are currently based on either photon counting or signal averaging. High information-density data acquisition (up to 3.2 GB/s of raw data) enables the smooth transition between the two modalities on a pixel-by-pixel basis and the ultimate writing of much smaller files (few kB/s). Modeling of the PMT response allows extraction of key sensor parameters from the histogram of voltage peak-heights. Applications in second harmonic generation (SHG) microscopy are described demonstrating S/N approaching the shot-noise limit of the detector over large dynamic ranges.

Citation Download Citation

David J. Kissick, Ryan D. Muir, Shane Z. Sullivan, Robert A. Oglesbee, and Garth J Simpson "Real-time dynamic range and signal to noise enhancement in beam-scanning microscopy by integration of sensor characteristics, data acquisition hardware, and statistical methods", Proc. SPIE 8657, Computational Imaging XI, 86570E (26 February 2013); https://doi.org/10.1117/12.2008607

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