Double-kinetic time-resolved measurement of non-equilibrium fluorescence lifetime with time sequence of nanosecond-scale: a novel technique based on single photon counting

2001 ◽

Author(s):

Mike Bruce ◽

Rama R. Goruganthu ◽

Shawn McBride ◽

David Bethke ◽

J.M. Chin

Keyword(s):

Single Photon ◽

Single Point ◽

Photon Counting ◽

Ring Oscillator ◽

Time Resolved ◽

Single Photon Counting ◽

Photo Diode ◽

Avalanche Photo Diode ◽

Photo Multiplier Tube ◽

Channel Plate

Abstract For time resolved hot carrier emission from the backside, an alternate approach is demonstrated termed single point PICA. The single point approach records time resolved emission from an individual transistor using time-correlated-single-photon counting and an avalanche photo-diode. The avalanche photo-diode has a much higher quantum efficiency than micro-channel plate photo-multiplier tube based imaging cameras typically used in earlier approaches. The basic system is described and demonstrated from the backside on a ring oscillator circuit.

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Simple scanning near-infrared time-correlated single-photon counting instrument with a pulsed diode laser and avalanche photodiode (APD) for time-resolved measurements in scanning applications with a DNA sequencer

10.1117/12.347539 ◽

1999 ◽

Cited By ~ 2

Author(s):

Yuling Zhang ◽

Steven A. Soper ◽

Lyle R. Middendorf ◽

John A. Wrum ◽

Rainer Erdmann ◽

...

Keyword(s):

Diode Laser ◽

Near Infrared ◽

Single Photon ◽

Photon Counting ◽

Avalanche Photodiode ◽

Time Resolved ◽

Single Photon Counting ◽

Time Resolved Measurements ◽

Dna Sequencer

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Optical Estimation of Absolute Membrane Potential Using One- and Two-Photon Fluorescence Lifetime Imaging Microscopy

10.1101/2021.02.16.431491 ◽

2021 ◽

Author(s):

Julia R. Lazzari-Dean ◽

Evan W. Miller

Keyword(s):

Membrane Potential ◽

Fluorescence Lifetime ◽

Single Photon ◽

Photon Counting ◽

Fluorescence Lifetime Imaging ◽

Single Photon Counting ◽

Two Photon ◽

Lifetime Imaging ◽

Wide Range ◽

Relative Measurements

AbstractBackgroundMembrane potential (Vmem) exerts physiological influence across a wide range of time and space scales. To study Vmem in these diverse contexts, it is essential to accurately record absolute values of Vmem, rather than solely relative measurements.Materials & MethodsWe use fluorescence lifetime imaging of a small molecule voltage sensitive dye (VF2.1.Cl) to estimate mV values of absolute membrane potential.ResultsWe test the consistency of VF2.1.Cl lifetime measurements performed on different single photon counting instruments and find that they are in striking agreement (differences of <0.5 ps/mV in the slope and <50 ps in the y-intercept). We also demonstrate that VF2.1.Cl lifetime reports absolute Vmem under two-photon (2P) illumination with better than 20 mV of Vmem resolution, a nearly 10-fold improvement over other lifetime-based methods.ConclusionsWe demonstrate that VF-FLIM is a robust and portable metric for Vmem across imaging platforms and under both one-photon and two-photon illumination. This work is a critical foundation for application of VF-FLIM to record absolute membrane potential signals in thick tissue.

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