Fluorescence lifetime imaging of high-speed particles with single-photon image sensors

2019 ◽  
Author(s):  
Istvan Gyongy ◽  
Andrew Green ◽  
Sam W. Hutchings ◽  
Amy Davies ◽  
Neale Dutton ◽  
...  
Keyword(s):  
Fluorescence Lifetime ◽  
High Speed ◽  
Single Photon ◽  
Image Sensors ◽  
Fluorescence Lifetime Imaging ◽  
Lifetime Imaging

scholarly journals Video-rate fluorescence lifetime imaging camera with CMOS single-photon avalanche diode arrays and high-speed imaging algorithm

2011 ◽  
Vol 16 (9) ◽  
pp. 096012 ◽  
Author(s):  
David D.-U. Li ◽  
Jochen Arlt ◽  
David Tyndall ◽  
Richard Walker ◽  
Justin Richardson ◽  
...  
Keyword(s):  
Fluorescence Lifetime ◽  
High Speed ◽  
Single Photon ◽  
Fluorescence Lifetime Imaging ◽  
High Speed Imaging ◽  
Imaging Algorithm ◽  
Avalanche Diode ◽  
Lifetime Imaging ◽  
Imaging Camera

scholarly journals Wide-field Fluorescence Lifetime Imaging Microscopy with a High-Speed Mega-pixel SPAD Camera

2020 ◽  
Author(s):  
V. Zickus ◽  
M.-L. Wu ◽  
K. Morimoto ◽  
V. Kapitany ◽  
A. Fatima ◽  
...  
Keyword(s):  
Fluorescence Lifetime ◽  
High Speed ◽  
Single Photon ◽  
Fluorescent Proteins ◽  
Cell Metabolism ◽  
Fluorescence Lifetime Imaging Microscopy ◽  
Wide Field ◽  
Fluorescence Lifetime Imaging ◽  
Protein Activity ◽  
Lifetime Imaging

Fluorescence lifetime imaging microscopy (FLIM) is a key technology that provides direct insight into cell metabolism, cell dynamics and protein activity. However, determining the lifetimes of different fluorescent proteins requires the detection of a relatively large number of photons, hence slowing down total acquisition times. Moreover, there are many cases, for example in studies of cell collectives, where wide-field imaging is desired. We report scan-less wide-field FLIM based on a 0.5 Megapixel resolution, time-gated Single Photon Avalanche Diode (SPAD) camera, with acquisition rates up to 1 Hz. Fluorescence lifetime estimation is performed via a pre-trained artificial neural network with 1000-fold improvement in processing times compared to standard least squares fitting techniques. We utilised our system to image HT1080 – human fibrosarcoma cell line as well as Convallaria. The results show promise for real-time FLIM and a viable route towards multi-megapixel fluorescence lifetime images, with a proof-of-principle mosaic image shown with 3.6 megapixels.

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 High-speed imaging of transient metabolic dynamics using two-photon fluorescence lifetime imaging microscopy

Optica ◽  
2018 ◽  
Vol 5 (10) ◽  
pp. 1290 ◽  
Author(s):  
Andrew J. Bower ◽  
Joanne Li ◽  
Eric J. Chaney ◽  
Marina Marjanovic ◽  
Darold R. Spillman ◽  
...  
Keyword(s):  
Fluorescence Lifetime ◽  
High Speed ◽  
Fluorescence Lifetime Imaging Microscopy ◽  
Fluorescence Lifetime Imaging ◽  
High Speed Imaging ◽  
Two Photon ◽  
Lifetime Imaging ◽  
Two Photon Fluorescence ◽  
Photon Fluorescence

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
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