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Related Concept Videos

Super-resolution Fluorescence Microscopy01:37

Super-resolution Fluorescence Microscopy

Super-resolution fluorescence microscopy (SRFM) provides a better resolution than conventional fluorescence microscopy by reducing the point spread function (PSF). PSF is the light intensity distribution from a point that causes it to appear blurred. Due to PSF, each fluorescing point appears bigger than its actual size, and it is the PSF interference of nearby fluorophores that causes the blurred image. Various approaches to achieving higher resolution through SRFM have recently been developed.

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    This study introduces an Active Quenching Circuit (AQC) to overcome the pile-up effect in Time-Correlated Single Photon Counting (TCSPC). The novel circuit enables distortionless measurements even at high photon rates, enhancing sensitivity in applications like FLIM.

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    Area of Science:

    • Instrumentation and Measurement Science
    • Photonics and Optical Engineering

    Background:

    • Time-Correlated Single Photon Counting (TCSPC) is crucial for low-light detection in fluorescence lifetime imaging microscopy (FLIM) and quantum optics.
    • The pile-up effect significantly distorts TCSPC measurements at high photon detection rates, limiting performance.
    • Existing solutions for pile-up often involve complex post-processing or multichannel systems, complicating setups.

    Purpose of the Study:

    • To develop a novel Active Quenching Circuit (AQC) to mitigate the pile-up effect in TCSPC.
    • To achieve distortionless TCSPC histograms at high illumination conditions by precisely controlling photodetector dead time.
    • To ensure robust and precise dead time tuning resistant to environmental variations.

    Main Methods:

    • Development of an Active Quenching Circuit (AQC) using high-voltage 150 nm technology.
    • Implementation of Process, Voltage, and Temperature (PVT) compensation for stable dead time control.
    • Experimental validation using fluorescence measurements under elevated count-rate conditions.

    Main Results:

    • The AQC achieved a dead-time resolution of 50 ps, suitable for laser frequencies from 20 to 100 MHz.
    • Demonstrated close-to-ideal linearity in dead-time control across varying conditions.
    • Experimental results showed a distortion as low as 0.43% under high count-rate conditions.

    Conclusions:

    • The developed AQC effectively overcomes the pile-up limitation in TCSPC.
    • This advancement enables more accurate and sensitive measurements in demanding optical applications.
    • The circuit offers a robust and precise solution for time-resolved experiments.