scholarly journals Single-Photon, Time-Gated, Phasor-Based Fluorescence Lifetime Imaging through Highly Scattering Medium

ACS Photonics ◽  
2019 ◽  
Vol 7 (1) ◽  
pp. 68-79 ◽  
Author(s):  
Rinat Ankri ◽  
Arkaprabha Basu ◽  
Arin Can Ulku ◽  
Claudio Bruschini ◽  
Edoardo Charbon ◽  
...  
Keyword(s):  
Fluorescence Lifetime ◽  
Single Photon ◽  
Fluorescence Lifetime Imaging ◽  
Scattering Medium ◽  
Lifetime Imaging ◽  
Highly Scattering Medium

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

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.

scholarly journals Real-time multispectral fluorescence lifetime imaging using Single Photon Avalanche Diode arrays

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
João L. Lagarto ◽  
Federica Villa ◽  
Simone Tisa ◽  
Franco Zappa ◽  
Vladislav Shcheslavskiy ◽  
...  
Keyword(s):  
Real Time ◽  
Fluorescence Lifetime ◽  
Single Photon ◽  
Fluorescence Lifetime Imaging ◽  
Avalanche Diode ◽  
Lifetime Imaging

Fluorescence lifetime imaging with near-infrared dyes

2015 ◽  
Vol 4 (1) ◽  
Author(s):  
Wolfgang Becker ◽  
Vladislav Shcheslavskiy
Keyword(s):  
Fluorescence Lifetime ◽  
Laser Scanning ◽  
Near Infrared ◽  
Single Photon ◽  
Optical Tomography ◽  
Small Animal Imaging ◽  
Supplementary Information ◽  
Fluorescence Lifetime Imaging ◽  
Lifetime Imaging ◽  
Nir Dyes

AbstractNear-infrared (NIR) dyes are used as fluorescence markers in small animal imaging and in diffuse optical tomography. In these applications it is important to know whether the dyes bind to proteins or to other tissue constituents, and whether their fluorescence lifetimes depend on the targets they bind to. Unfortunately, neither the optical beam paths nor the detectors of commonly used in confocal and multiphoton laser scanning microscopes (LSMs) directly allow for excitation and detection of NIR fluorescence. This paper presents three ways of adapting existing LSMs with time-correlated single photon counting (TCSPC) fluorescence lifetime imaging (FLIM) systems for NIR FLIM: 1) confocal systems with wideband beamsplitters and diode laser excitation, 2) confocal systems with wideband beamsplitters and one-photon excitation by titanium-sapphire lasers, and 3) two-photon systems with optical parametric oscillator (OPO) excitation and non-descanned detection. A number of NIR dyes are tested in biological tissue. All of them show clear lifetime changes depending on the tissue structures they are bound to. We therefore believe that NIR FLIM can deliver supplementary information about the tissue composition and on local biochemical parameters.

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