scholarly journals Full-field fluorescence lifetime dual-comb microscopy using spectral mapping and frequency multiplexing of dual-comb optical beats

2021 ◽  
Vol 7 (1) ◽  
pp. eabd2102
Author(s):  
T. Mizuno ◽  
E. Hase ◽  
T. Minamikawa ◽  
Y. Tokizane ◽  
R. Oe ◽  
...  
Keyword(s):  
Fluorescence Imaging ◽  
Fluorescence Lifetime ◽  
Detection Efficiency ◽  
Fluorescence Lifetime Imaging ◽  
Spectral Mapping ◽  
Vast Number ◽  
Full Field ◽  
Mechanical Scanning ◽  
Frequency Multiplexing ◽  
Quantitative Fluorescence

Fluorescence lifetime imaging microscopy (FLIM) is a powerful tool for quantitative fluorescence imaging because fluorescence lifetime is independent of concentration of fluorescent molecules or excitation/detection efficiency and is robust to photobleaching. However, since most FLIMs are based on point-to-point measurements, mechanical scanning of a focal spot is needed for forming an image, which hampers rapid imaging. Here, we demonstrate scan-less full-field FLIM based on a one-to-one correspondence between two-dimensional (2D) image pixels and frequency-multiplexed radio frequency (RF) signals. A vast number of dual-comb optical beats between dual optical frequency combs are effectively adopted for 2D spectral mapping and high-density frequency multiplexing in the RF region. Bimodal images of fluorescence amplitude and lifetime are obtained with high quantitativeness from amplitude and phase spectra of fluorescence RF comb modes without the need for mechanical scanning. The parallelized FLIM will be useful for rapid quantitative fluorescence imaging in life science.

scholarly journals Timing and Operating Mode Design for Time-Gated Fluorescence Lifetime Imaging Microscopy

2013 ◽  
Vol 2013 ◽  
pp. 1-5 ◽  
Author(s):  
Chao Liu ◽  
Xinwei Wang ◽  
Yan Zhou ◽  
Yuliang Liu
Keyword(s):  
High Resolution ◽  
Fluorescence Imaging ◽  
Fluorescence Lifetime ◽  
Imaging Techniques ◽  
Absolute Measurement ◽  
Operating Mode ◽  
Least Square ◽  
Fluorescence Lifetime Imaging Microscopy ◽  
Fluorescence Lifetime Imaging ◽  
Lifetime Imaging

Steady-state fluorence imaging and time-resolved fluorescence imaging are two important areas in fluorescence imaging research. Fluorescence lifetime imaging is an absolute measurement method which is independent of excitation laser intensity, fluorophore concentration, and photobleaching compared to fluorescence intensity imaging techniques. Time-gated fluorescence lifetime imaging microscopy (FLIM) can provide high resolution and high imaging frame during mature FLIM methods. An abstract time-gated FLIM model was given, and important temporal parameters are shown as well. Aiming at different applications of steady and transient fluorescence processes, two different operation modes, timing and lifetime computing algorithm are designed. High resolution and high frame can be achieved by one-excitation one-sampling mode and least square algorithm for steady imaging applications. Correspondingly, one-excitation two-sampling mode and rapid lifetime determination algorithm contribute to transient fluorescence situations.

scholarly journals Deep Learning Enhanced Hyperspectral Fluorescence Lifetime Imaging

2020 ◽  
Author(s):  
Marien Ochoa ◽  
Alena Rudkouskaya ◽  
Ruoyang Yao ◽  
Pingkun Yan ◽  
Margarida Barroso ◽  
...  
Keyword(s):  
Deep Learning ◽  
Fluorescence Lifetime ◽  
Detection Efficiency ◽  
Optical Data ◽  
Fluorescence Lifetime Imaging ◽  
Her2 Positive ◽  
Lifetime Imaging ◽  
Intracellular Drug Delivery ◽  
Single Pixel

Acquiring dense high-dimensional optical data in biological applications remains a challenge due to the very low levels of light typically encountered. Single pixel imaging methodologies enable improved detection efficiency in such conditions but are still limited by relatively slow acquisition times. Here, we propose a Deep Learning framework, NetFLICS-CR, which enables fast hyperspectral lifetime imaging for in vivo applications at enhanced resolution, acquisition and processing speeds, without the need of experimental training datasets. NetFLICS-CR reconstructs intensity and lifetime images at 128×128 pixels over 16 spectral channels while reducing the current acquisition times from ∼2.5 hours at 50% compression to ∼3 minutes at 99% compression when using a single-pixel Hyperspectral Macroscopic Fluorescence Lifetime Imaging (HMFLI) system. The potential of the technique is demonstrated in silico, in vitro and in vivo through the monitoring of receptor-ligand interactions in mice liver and bladder and further imaging of intracellular drug delivery of the clinical drug Trastuzumab in live animals bearing HER2-positive breast tumor xenografts.

scholarly journals Multiphoton fluorescence lifetime imaging microscopy (FLIM) and super-resolution fluorescence imaging with a supramolecular biopolymer for the controlled tagging of polysaccharides

Nanoscale ◽  
2019 ◽  
Vol 11 (19) ◽  
pp. 9498-9507 ◽  
Author(s):  
Haobo Ge ◽  
Fernando Cortezon-Tamarit ◽  
Hui-Chen Wang ◽  
Adam C. Sedgwick ◽  
Rory L. Arrowsmith ◽  
...  
Keyword(s):  
Fluorescence Imaging ◽  
Fluorescence Lifetime ◽  
Super Resolution ◽  
Fluorescence Lifetime Imaging Microscopy ◽  
Fluorescence Lifetime Imaging ◽  
Boronate Ester ◽  
Lifetime Imaging ◽  
In Cells

A new coumarin-appended boronate ester for fluorogenic imaging which binds polysaccharides in solution and in cells.

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