A wide-field time-domain fluorescence lifetime imaging microscope with optical sectioning

2002 ◽  
Vol 73 (4) ◽  
pp. 1898-1907 ◽  
Author(s):  
S. E. D. Webb ◽  
Y. Gu ◽  
S. Lévêque-Fort ◽  
J. Siegel ◽  
M. J. Cole ◽  
...  
Keyword(s):  
Fluorescence Lifetime ◽  
Time Domain ◽  
Wide Field ◽  
Fluorescence Lifetime Imaging ◽  
Optical Sectioning ◽  
Lifetime Imaging

scholarly journals Wide-field fluorescence lifetime imaging with optical sectioning and spectral resolution applied to biological samples

2002 ◽  
Vol 49 (5-6) ◽  
pp. 985-995 ◽  
Author(s):  
D. S. Elson ◽  
J. Siegel ◽  
S. E. D. Webb ◽  
S. Lévêquefort ◽  
D. Parsons-Karavassilis ◽  
...  
Keyword(s):  
Fluorescence Lifetime ◽  
Spectral Resolution ◽  
Biological Samples ◽  
Wide Field ◽  
Fluorescence Lifetime Imaging ◽  
Optical Sectioning ◽  
Lifetime Imaging

Time-domain whole-field fluorescence lifetime imaging with optical sectioning

2001 ◽  
Vol 203 (3) ◽  
pp. 246-257 ◽  
Author(s):  
M. J. Cole ◽  
J. Siegel ◽  
S. E. D. Webb ◽  
R. Jones ◽  
K. Dowling ◽  
...  
Keyword(s):  
Fluorescence Lifetime ◽  
Time Domain ◽  
Fluorescence Lifetime Imaging ◽  
Optical Sectioning ◽  
Lifetime Imaging

scholarly journals Macroscopic fluorescence-lifetime imaging of NADH and protoporphyrin IX improves the detection and grading of 5-aminolevulinic acid-stained brain tumors

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Mikael T. Erkkilä ◽  
David Reichert ◽  
Johanna Gesperger ◽  
Barbara Kiesel ◽  
Thomas Roetzer ◽  
...  
Keyword(s):  
Brain Tumor ◽  
Brain Tumors ◽  
Fluorescence Lifetime ◽  
Tumor Tissue ◽  
Aminolevulinic Acid ◽  
Protoporphyrin Ix ◽  
Low Grade ◽  
Wide Field ◽  
Fluorescence Lifetime Imaging ◽  
Lifetime Imaging

AbstractMaximal safe tumor resection remains the key prognostic factor for improved prognosis in brain tumor patients. Despite 5-aminolevulinic acid-based fluorescence guidance the neurosurgeon is, however, not able to visualize most low-grade gliomas (LGG) and infiltration zone of high-grade gliomas (HGG). To overcome the need for a more sensitive visualization, we investigated the potential of macroscopic, wide-field fluorescence lifetime imaging of nicotinamide adenine dinucleotide (NADH) and protoporphyrin IX (PPIX) in selected human brain tumors. For future intraoperative use, the imaging system offered a square field of view of 11 mm at 250 mm free working distance. We performed imaging of tumor tissue ex vivo, including LGG and HGG as well as brain metastases obtained from 21 patients undergoing fluorescence-guided surgery. Half of all samples showed visible fluorescence during surgery, which was associated with significant increase in PPIX fluorescence lifetime. While the PPIX lifetime was significantly different between specific tumor tissue types, the NADH lifetimes did not differ significantly among them. However, mainly necrotic areas exhibited significantly lower NADH lifetimes compared to compact tumor in HGG. Our pilot study indicates that combined fluorescence lifetime imaging of NADH/PPIX represents a sensitive tool to visualize brain tumor tissue not detectable with conventional 5-ALA fluorescence.

scholarly journals Optically sectioned wide-field fluorescence lifetime imaging microscopy enabled by structured illumination

2017 ◽  
Vol 8 (3) ◽  
pp. 1455 ◽  
Author(s):  
Taylor Hinsdale ◽  
Cory Olsovsky ◽  
Jose J. Rico-Jimenez ◽  
Kristen C. Maitland ◽  
Javier A. Jo ◽  
...  
Keyword(s):  
Fluorescence Lifetime ◽  
Fluorescence Lifetime Imaging Microscopy ◽  
Wide Field ◽  
Fluorescence Lifetime Imaging ◽  
Structured Illumination ◽  
Lifetime Imaging

High-speed wide-field time-gated endoscopic fluorescence-lifetime imaging

2004 ◽  
Vol 29 (19) ◽  
pp. 2249 ◽  
Author(s):  
J. Requejo-Isidro ◽  
J. McGinty ◽  
I. Munro ◽  
D. S. Elson ◽  
N. P. Galletly ◽  
...  
Keyword(s):  
Fluorescence Lifetime ◽  
High Speed ◽  
Wide Field ◽  
Fluorescence Lifetime Imaging ◽  
Lifetime Imaging

scholarly journals In Vivo Time Domain Optical Imaging of Renal Ischemia-Reperfusion Injury: Discrimination Based on Fluorescence Lifetime

2007 ◽  
Vol 6 (5) ◽  
pp. 7290.2007.00030 ◽  
Author(s):  
Abedelnasser Abulrob ◽  
Eric Brunette ◽  
Jacqueline Slinn ◽  
Ewa Baumann ◽  
Danica Stanimirovic
Keyword(s):  
Optical Imaging ◽  
Fluorescence Lifetime ◽  
Fluorescence Intensity ◽  
Time Domain ◽  
Ex Vivo ◽  
Renal Ischemia ◽  
Fluorescence Lifetime Imaging ◽  
Lifetime Imaging ◽  
Lifetime Analysis

Fluorescence lifetime is an intrinsic parameter of the fluorescent probe, independent of the probe concentration but sensitive to changes in the surrounding microenvironment. Therefore, fluorescence lifetime imaging could potentially be applied to in vivo diagnostic assessment of changes in the tissue microenvironment caused by disease, such as ischemia. The aim of this study was to evaluate the utility of noninvasive fluorescence lifetime imaging in distinguishing between normal and ischemic kidney tissue in vivo. Mice were subjected to 60-minute unilateral kidney ischemia followed by 6-hour reperfusion. Animals were then injected with the near-infrared fluorescence probe Cy5.5 or saline and imaged using a time-domain small-animal optical imaging system. Both fluorescence intensity and lifetime were acquired. The fluorescence intensity of Cy5.5 was clearly reduced in the ischemic compared with the contralateral kidney, and the fluorescence lifetime of Cy5.5 was not detected in the ischemic kidney, suggesting reduced kidney clearance. Interestingly, the two-component lifetime analysis of endogenous fluorescence at 700 nm distinguished renal ischemia in vivo without the need for Cy5.5 injection for contrast enhancement. The average fluorescence lifetime of endogenous tissue fluorophores was a sensitive indicator of kidney ischemia ex vivo. The study suggests that fluorescence lifetime analysis of endogenous tissue fluorophores could be used to discriminate ischemic or necrotic tissues by noninvasive in vivo or ex vivo organ imaging.

scholarly journals Single Photon, Time-Gated, Phasor-based Fluorescence Lifetime Imaging Through Highly Scattering Medium

2019 ◽  
Author(s):  
Rinat Ankri ◽  
Arkaprabha Basu ◽  
Arin Can Ulku ◽  
Claudio Bruschini ◽  
Edoardo Charbon ◽  
...  
Keyword(s):  
Fluorescence Lifetime ◽  
Fluorescence Intensity ◽  
Single Photon ◽  
Acquisition Time ◽  
Fluorescence Signal ◽  
Wide Field ◽  
Fluorescence Lifetime Imaging ◽  
Short Acquisition Time ◽  
Lifetime Imaging

AbstractFluorescence lifetime imaging (FLI) is a powerful tool for in vitro and non-invasive in vivo biomolecular and cellular investigations. Fluorescence lifetime is an intrinsic characteristic of any fluorescent dye which, to some extent, does not depend on excitation intensity and signal level. However, when used in vivo with visible wavelength emitting fluorophores, FLI is complicated by (i) light scattering as well as absorption by tissues, which significantly reduces fluorescence intensity, (ii) tissue autofluorescence (AF), which decreases the signal to noise ratio and (iii) broadening of the decay signal, which can result in incorrect lifetime estimation. Here, we report the use of a large-frame time-gated single-photon avalanche diode (SPAD) imager, SwissSPAD2, with a very short acquisition time (in the milliseconds range) and a wide-field microscopy format. We use the phasor approach to convert each pixel’s data into its local lifetime. The phasor transformation provides a simple and fast visual method for lifetime imaging and is particularly suitable for in vivo FLI which suffers from deformation of the fluorescence decay, and makes lifetime extraction by standard fitting challenging. We show, for single dyes, that the phasor cloud distribution (of pixels) increases with decay broadening due to scattering and decreasing fluorescence intensity. Yet, as long as the fluorescence signal is higher than the tissue-like phantom AF, a distinct lifetime can still be clearly identified with an appropriate background correction. Lastly, we demonstrate the detection of few hundred thousand A459 cells expressing the fluorescent protein mCyRFP1 through highly scattering phantom layers, despite significant scattering and the presence of the phantom AF.

scholarly journals Data Processing for Time-Domain Fluorescence Lifetime Imaging Microscopy

2013 ◽  
Vol 19 (S2) ◽  
pp. 758-759
Author(s):  
P.A. Young ◽  
A. Grislis ◽  
P.R. Barber ◽  
P.J. Keely ◽  
K.W. Eliceiri
Keyword(s):  
Data Processing ◽  
Fluorescence Lifetime ◽  
Time Domain ◽  
Fluorescence Lifetime Imaging Microscopy ◽  
Fluorescence Lifetime Imaging ◽  
Lifetime Imaging

Extended abstract of a paper presented at Microscopy and Microanalysis 2013 in Indianapolis, Indiana, USA, August 4 – August 8, 2013.

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