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Time-gated luminescence microscopy.

Russell E Connally¹, James A Piper

¹Centre for Laser Applications, Department of Physics, Division of Information and Communication Sciences, Macquarie University, Sydney, NSW 2109, Australia. rconnall@ics.mq.edu.au

Annals of the New York Academy of Sciences

|July 4, 2008

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A new time-gated luminescence (TGL) microscope enhances signal-to-noise ratio by 14x for clearer imaging. This advanced microscopy technique suppresses short-lived fluorescence, improving detection of specific signals.

Area of Science:

Biomedical Optics
Microscopy Technology
Analytical Chemistry

Background:

Autofluorescence from biological samples can obscure target signals.
Short-lived fluorescence complicates imaging, reducing signal-to-noise ratio.
Existing microscopy techniques may struggle with autofluorescence suppression.

Purpose of the Study:

To introduce and evaluate a novel all-solid-state, time-gated luminescence (TGL) microscope.
To demonstrate the TGL microscope's capability in suppressing short-lived fluorescence.
To assess the sensitivity and signal-to-noise improvement of the TGL system.

Main Methods:

Utilized pulsed UV excitation (365 nm) with a UV-LED source on an Olympus BX51 microscope.
Employed an electron multiplying charge-coupled-device (CCD) camera in delayed luminescence mode.

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Last Updated: Jul 4, 2026

Luminescence Lifetime Imaging of O2 with a Frequency-Domain-Based Camera System

Luminescence Lifetime Imaging of O₂ with a Frequency-Domain-Based Camera System

Published on: December 16, 2019

Fluorescence Lifetime Imaging of Molecular Rotors in Living Cells

Fluorescence Lifetime Imaging of Molecular Rotors in Living Cells

Published on: February 9, 2012

Luminescence Resonance Energy Transfer to Study Conformational Changes in Membrane Proteins Expressed in Mammalian Cells

Luminescence Resonance Energy Transfer to Study Conformational Changes in Membrane Proteins Expressed in Mammalian Cells

Published on: September 16, 2014

Analyzed autofluorescent algal samples spiked with europium beads and immunofluorescently labeled Giardia cysts.

Main Results:

Achieved a 14-fold increase in signal-to-noise ratio within a 33 ms capture cycle.
Demonstrated successful suppression of short-lived fluorescence by triggering the camera 3 microseconds after excitation.
Showcased the instrument's ability to switch between prompt epifluorescence and TGL modes.

Conclusions:

The novel all-solid-state TGL microscope offers significant improvements in imaging sensitivity and clarity.
This TGL microscopy approach effectively suppresses autofluorescence and short-lived signals.
The instrument provides a versatile platform for enhanced biological sample analysis.