Fluorescence lifetime estimation of multiple near-infrared dyes in mice

2008 ◽  
Author(s):  
Guobin Ma ◽  
Simon Fortier ◽  
Muriel Jean-Jacques ◽  
Niculae Mincu ◽  
Frederic Leblond ◽  
...  
Keyword(s):  
Fluorescence Lifetime ◽  
Near Infrared ◽  
Lifetime Estimation

scholarly journals Optimization of Advanced Live-Cell Imaging through Red/Near-Infrared Dye Labeling and Fluorescence Lifetime-Based Strategies

2021 ◽  
Vol 22 (20) ◽  
pp. 11092
Author(s):  
Magalie Bénard ◽  
Damien Schapman ◽  
Christophe Chamot ◽  
Fatéméh Dubois ◽  
Guénaëlle Levallet ◽  
...  
Keyword(s):  
Fluorescence Lifetime ◽  
Live Cell Imaging ◽  
Near Infrared ◽  
Cell Imaging ◽  
Light Exposure ◽  
Photon Counting ◽  
Live Cell ◽  
Stimulated Emission ◽  
Plot Analysis ◽  
Phasor Plot

Fluorescence microscopy is essential for a detailed understanding of cellular processes; however, live-cell preservation during imaging is a matter of debate. In this study, we proposed a guide to optimize advanced light microscopy approaches by reducing light exposure through fluorescence lifetime (τ) exploitation of red/near-infrared dyes. Firstly, we characterized key instrumental elements which revealed that red/near-infrared laser lines with an 86x (Numerical Aperture (NA) = 1.2, water immersion) objective allowed high transmission of fluorescence signals, low irradiance and super-resolution. As a combination of two technologies, i.e., vacuum tubes (e.g., photomultiplier) and semiconductor microelectronics (e.g., avalanche photodiode), type S, X and R of hybrid detectors (HyD-S, HyD-X and HyD-R) were particularly adapted for red/near-infrared photon counting and τ separation. Secondly, we tested and compared lifetime-based imaging including coarse τ separation for confocal microscopy, fitting and phasor plot analysis for fluorescence lifetime microscopy (FLIM), and lifetimes weighting for enhanced stimulated emission depletion (STED) nanoscopy, in light of red/near-infrared multiplexing. Mainly, we showed that the choice of appropriate imaging approach may depend on fluorochrome number, together with their spectral/lifetime characteristics and STED compatibility. Photon-counting mode and sensitivity of HyDs together with phasor plot analysis of fluorescence lifetimes enabled the flexible and fast imaging of multi-labeled living H28 cells. Therefore, a combination of red/near-infrared dyes labeling with lifetime-based strategies offers new perspectives for live-cell imaging by enhancing sample preservation through acquisition time and light exposure reduction.

scholarly journals Spectroscopically Well-Characterized RGD Optical Probe as a Prerequisite for Lifetime-Gated Tumor Imaging

2011 ◽  
Vol 10 (6) ◽  
pp. 7290.2011.00018 ◽  
Author(s):  
Julia Eva Mathejczyk ◽  
Jutta Pauli ◽  
Christian Dullin ◽  
Joanna Napp ◽  
Lutz-F. Tietze ◽  
...  
Keyword(s):  
Fluorescence Lifetime ◽  
Nude Mice ◽  
Near Infrared ◽  
Tumor Imaging ◽  
Specific Binding ◽  
Optical Probe ◽  
Detection Sensitivity ◽  
Rgd Peptides ◽  
Emissive Probes

Labeling of RGD peptides with near-infrared fluorophores yields optical probes for noninvasive imaging of tumors overexpressing αvβ3 integrins. An important prerequisite for optimum detection sensitivity in vivo is strongly absorbing and highly emissive probes with a known fluorescence lifetime. The RGD-Cy5.5 optical probe was derived by coupling Cy5.5 to a cyclic arginine–glycine–aspartic acid–d-phenylalanine–lysine (RGDfK) peptide via an aminohexanoic acid spacer. Spectroscopic properties of the probe were studied in different matrices in comparison to Cy5.5. For in vivo imaging, human glioblastoma cells were subcutaneously implanted into nude mice, and in vivo fluorescence intensity and lifetime were measured. The fluorescence quantum yield and lifetime of Cy5.5 were found to be barely affected on RGD conjugation but dramatically changed in the presence of proteins. By time domain fluorescence imaging, we demonstrated specific binding of RGD-Cy5.5 to glioblastoma xenografts in nude mice. Discrimination of unspecific fluorescence by lifetime-gated analysis further enhanced the detection sensitivity of RGD-Cy5.5-derived signals. We characterized RGD-Cy5.5 as a strongly emissive and stable probe adequate for selective targeting of αvβ3 integrins. The specificity and thus the overall detection sensitivity in vivo were optimized with lifetime gating, based on the previous determination of the probes fluorescence lifetime under application-relevant conditions.

In vivo imaging with near-infrared fluorescence lifetime contrast

2009 ◽  
Author(s):  
Walter J. Akers ◽  
Mikhail Y. Berezin ◽  
Hyeran Lee ◽  
Samuel Achilefu
Keyword(s):  
Fluorescence Lifetime ◽  
In Vivo Imaging ◽  
Near Infrared ◽  
Near Infrared Fluorescence

scholarly journals Fluorescence lifetime imaging of upper gastrointestinal pH in vivo with a lanthanide based near-infrared τ probe

2019 ◽  
Vol 10 (15) ◽  
pp. 4227-4235 ◽  
Author(s):  
Yingying Ning ◽  
Shengming Cheng ◽  
Jing-Xiang Wang ◽  
Yi-Wei Liu ◽  
Wei Feng ◽  
...  
Keyword(s):  
Fluorescence Lifetime ◽  
Near Infrared ◽  
Lanthanide Complex ◽  
Fluorescence Lifetime Imaging ◽  
Ph Responsive ◽  
Gastrointestinal Ph ◽  
Lifetime Imaging ◽  
Upper Gastrointestinal

Lanthanide complex was successfully applied in the design of pH-responsive NIR τ probe for quantitative in vivo imaging.

Fluorescence lifetime estimation using a dynamic vision sensor

2017 ◽  
Author(s):  
Fengqiang Li ◽  
Nathan Matsuda ◽  
Marc Walton ◽  
Oliver Cossairt
Keyword(s):  
Fluorescence Lifetime ◽  
Vision Sensor ◽  
Lifetime Estimation ◽  
Dynamic Vision

scholarly journals Fluorescence lifetime estimation method for incomplete decay

2015 ◽  
Vol 51 (21) ◽  
pp. 1666-1668 ◽  
Author(s):  
Hongqi Yu ◽  
David Day‐Uei Li
Keyword(s):  
Fluorescence Lifetime ◽  
Estimation Method ◽  
Lifetime Estimation

scholarly journals Substrate-Based Near-Infrared Imaging Sensors Enable Fluorescence Lifetime Contrast via Built-in Dynamic Fluorescence Quenching Elements

ACS Sensors ◽  
2016 ◽  
Vol 1 (4) ◽  
pp. 427-436 ◽  
Author(s):  
Anand T. N. Kumar ◽  
William L. Rice ◽  
Jessica C. López ◽  
Suresh Gupta ◽  
Craig J. Goergen ◽  
...  
Keyword(s):  
Fluorescence Quenching ◽  
Fluorescence Lifetime ◽  
Near Infrared ◽  
Infrared Imaging ◽  
Near Infrared Imaging ◽  
Imaging Sensors ◽  
Dynamic Fluorescence
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