All-Solid-State Diode-Pumped Fluorescence Lifetime Imaging System for Biomedicaine and Microscopy

1999 ◽  
Vol 5 (S2) ◽  
pp. 1066-1067
Author(s):  
M. J. Cole ◽  
K. Dowling ◽  
P. M. W. French ◽  
R. Jones ◽  
D. Parsons-Karavassilis ◽  
...  

The determination of fluorescence lifetime requires only relative measurements of intensity and so is especially useful for biomedical samples in which the heterogeneous nature of tissue and autofluorescence cause significant problems. Since fluorescence lifetime is dependent upon both radiative and non-radiative decay rates, it may be used to distinguish between different fluorophore molecules (with different radiative decay rates) and to monitor local environmental perturbations that affect the non-radiative decay rate. Fluorescence lifetime probes have been demonstrated for many biologically significant analytes including [O2], [Ca2+] and pH. Fluorescence lifetime imaging (FLIM) can be applied to almost any optical imaging modality, including microscopy and potentially to non-invasive optical biopsy. Fluorescence lifetime data may be acquired in the frequency or time domain. The recent development of user-friendly and relatively portable ultrafast laser technology and the availability of ultrafast gated optical image intensifiers (GOI’s) enable the development of potentially inexpensive time domain FLIM instruments that may be deployed outside specialist laser laboratories.

2021 ◽  
Author(s):  
Julia R. Lazzari-Dean ◽  
Evan W. Miller

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.


2005 ◽  
Vol 59 (9) ◽  
pp. 1174-1181 ◽  
Author(s):  
Daniela Comelli ◽  
Gianluca Valentini ◽  
Rinaldo Cubeddu ◽  
Lucia Toniolo

We developed a combined procedure for the analysis of works of art based on a portable system for fluorescence imaging integrated with analytical measurements on microsamples. The method allows us to localize and identify organic and inorganic compounds present on the surface of artworks. The fluorescence apparatus measures the temporal and spectral features of the fluorescence emission, excited by ultraviolet (UV) laser pulses. The kinetic of the emission is studied through a fluorescence lifetime imaging system, while an optical multichannel analyzer measures the fluorescence spectra of selected points. The chemical characterization of the compounds present on the artistic surfaces is then performed by means of analytical measurements on microsamples collected with the assistance of the fluorescence maps. The previous concepts have been successfully applied to study the contaminants on the surface of Michelangelo's David. The fluorescence analysis combined with Fourier transform infrared (FT-IR) measurements revealed the presence of beeswax, which permeates most of the statue surface, and calcium oxalate deposits mainly arranged in vertical patterns and related to rain washing.


2002 ◽  
Author(s):  
Michael Wahl ◽  
Hans-Juergen Rahn ◽  
Uwe Ortmann ◽  
Rainer Erdmann ◽  
Martin Boehmer ◽  
...  

2007 ◽  
Vol 6 (5) ◽  
pp. 7290.2007.00030 ◽  
Author(s):  
Abedelnasser Abulrob ◽  
Eric Brunette ◽  
Jacqueline Slinn ◽  
Ewa Baumann ◽  
Danica Stanimirovic

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.


1999 ◽  
Author(s):  
M. J. Cole ◽  
K. Dowling ◽  
R. Jones ◽  
D. Parsons-Karavassilis ◽  
P. M. W. French ◽  
...  

2015 ◽  
Vol 40 (14) ◽  
pp. 3296 ◽  
Author(s):  
Ximeng Y. Dow ◽  
Shane Z. Sullivan ◽  
Ryan D. Muir ◽  
Garth J. Simpson

2002 ◽  
Vol 73 (4) ◽  
pp. 1898-1907 ◽  
Author(s):  
S. E. D. Webb ◽  
Y. Gu ◽  
S. Lévêque-Fort ◽  
J. Siegel ◽  
M. J. Cole ◽  
...  

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