New Methods for Time-resolved Fluorescence Spectroscopy Data Analysis Based on the Laguerre Expansion Technique

2007 ◽  
Vol 46 (02) ◽  
pp. 206-211 ◽  
Author(s):  
L. Marcu ◽  
Q. Fang ◽  
T. Papaioannou ◽  
J. Qiao ◽  
M. Fishbein ◽  
...  

Summary Objectives : A new deconvolution method for the analysis of time-resolved laser-induced fluorescence spectroscopy (TR-LIFS) data is introduced and applied for tissue diagnosis. Method : The intrinsic TR-LIFS decays are expanded on a Laguerre basis, and the computed Laguerre expansion coefficients (LEC) are used to characterize the sample fluorescence emission. The method was applied for the diagnosis of atherosclerotic vulnerable plaques. Results : At a first stage, using a rabbit atherosclerotic model, 73 TR-LIFS in-vivo measurements from the normal and atherosclerotic aorta segments of eight rabbits were taken. The Laguerre deconvolution technique was able to accurately deconvolve the TR-LIFS measurements. More interesting, the LEC reflected the changes in the arterial biochemical composition and provided discrimination of lesions rich in macrophages/foamcells with high sensitivity (> 85%) and specificity (> 95%). At a second stage, 348 TR-LIFS measurements were obtained from the explanted carotid arteries of 30 patients. Lesions with significant inflammatory cells (macrophages/foam-cells and lymphocytes) were detected with high sensitivity (> 80%) and specificity (> 90%), using LEC-based classifiers. Conclusion : This study has demonstrated the potential of using TR-LIFS information by means of LEC for in- vivo tissue diagnosis, and specifically for detecting inflammation in atherosclerotic lesions, a key marker of plaque vulnerability.

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Md Imam Uddin ◽  
Tyler C. Kilburn ◽  
Sara Z. Jamal ◽  
Craig L. Duvall ◽  
John S. Penn

AbstractDiabetic retinopathy, retinopathy of prematurity and retinal vein occlusion are potentially blinding conditions largely due to their respective neovascular components. The development of real-time in vivo molecular imaging methods, to assess levels of retinal neovascularization (NV), would greatly benefit patients afflicted with these conditions. mRNA hybridization techniques offer a potential method to image retinal NV. The success of these techniques hinges on the selection of a target mRNA whose tissue levels and spatial expression patterns correlate closely with disease burden. Using a model of oxygen-induced retinopathy (OIR), we previously observed dramatic increases in retinal endoglin that localized to neovascular structures (NV), directly correlating with levels of neovascular pathology. Based on these findings, we have investigated Endoglin mRNA as a potential marker for imaging retinal NV in OIR mice. Also of critical importance, is the application of innovative technologies capable of detecting mRNAs in living systems with high sensitivity and specificity. To detect and visualize endoglin mRNA in OIR mice, we have designed and synthesized a novel imaging probe composed of short-hairpin anti-sense (AS) endoglin RNA coupled to a fluorophore and black hole quencher (AS-Eng shRNA). This assembly allows highly sensitive fluorescence emission upon hybridization of the AS-Eng shRNA to cellular endoglin mRNA. The AS-Eng shRNA is further conjugated to a diacyl-lipid (AS-Eng shRNA–lipid referred to as probe). The lipid moiety binds to serum albumin facilitating enhanced systemic circulation of the probe. OIR mice received intraperitoneal injections of AS-Eng shRNA–lipid. Ex vivo imaging of their retinas revealed specific endoglin mRNA dependent fluorescence superimposed on neovascular structures. Room air mice receiving AS-Eng shRNA–lipid and OIR mice receiving a non-sense control probe showed little fluorescence activity. In addition, we found that cells in neovascular lesions labelled with endoglin mRNA dependent fluorescence, co-labelled with the macrophage/microglia-associated marker IBA1. Others have shown that cells expressing macrophage/microglia markers associate with retinal neovascular structures in proportion to disease burden. Hence we propose that our probe may be used to image and to estimate the levels of retinal neovascular disease in real-time in living systems.


2020 ◽  
Vol 10 (3) ◽  
pp. 178-188
Author(s):  
Bipin Rooj ◽  
Ankita Dutta ◽  
Debojyoti Mukherjee ◽  
Sahidul Islam ◽  
Ujjwal Mandal

Background: Understanding the interaction between different organic dyes and carbon quantum dots helps us to understand several photo physical processes like electron transfer, energy transfer, molecular sensing, drug delivery and dye degradation processes etc. Objective: The primary objective of this study is to whether the carbon quantum dots can act as an electron donor and can participate in the different photo physical processes. Methods: In this work, Carbon Quantum Dots (CQDLs) are synthesized in most economical and simple carbonization method where petals of Nelumbo nucifera L. are used as a carbon precursor. The synthesized CQDLs were characterized by using experimental techniques like UV−Vis absorption, FT-IR, Transmission Electron Microscopy (TEM), steadystate and time-resolved fluorescence spectroscopy. Results: The spectral analysis shows that the so synthesized CQDLs are spherical in shape and its diameter is around 4.2 nm. It shows the fluorescence emission maximum at 495 nm with a quantum yield of 4%. In this work the interaction between Carbon Quantum Dots (CQDLs) and an organic dye Malachite Green (MG) is studied using fluorescence spectroscopic technique under ambient pH condition (At pH 7). The quenching mechanism of CQDLs with MG was investigated using Stern-Volmer equation and time-resolved fluorescence lifetime studies. The results show that the dominant process of fluorescence quenching is attributed to Forster Resonance Energy Transfer (FRET) having a donor acceptor distance of 53 Å where CQDLs act as a donor and MG acts as an acceptor. Conclusion: This work has a consequence that CQDLs can be used as a donor species for different photo physical processes such as photovoltaic cell, dye sensitized solar cell, and also for antioxidant activity study.


2004 ◽  
Vol 92 (7) ◽  
Author(s):  
Christian M. Marquardt ◽  
P. J. Panak ◽  
C. Apostolidis ◽  
A. Morgenstern ◽  
C. Walther ◽  
...  

SummaryThis work focuses on time-resolved laser fluorescence spectroscopy (TRLFS) of Pa(IV) in aqueous solution. Excitation at 308 nm causes a fluorescence emission with a peak maximum at about 460 nm. Thereby, the position of the band´s maximum depends on the concentration, the type of the acid, and the pH value. Increasing complexation of the Pa(IV) ion leads to an increasing hypsochromic shift of the emission band up to 46 nm. In contrast to the band position the half-width (61.6±1.4 nm) and the lifetime (16±2 ns) of the fluorescence emission do not change significantly with changes in the chemical environment of the Pa(IV). The results of this work show that speciation of Pa(IV) can be performed even in aqueous solution by using TRLFS.


2020 ◽  
Vol 11 (1) ◽  
Author(s):  
Xiaochen Qiu ◽  
Qianwen Zhou ◽  
Xingjun Zhu ◽  
Zugen Wu ◽  
Wei Feng ◽  
...  

AbstractThe in vivo temperature monitoring of a microenvironment is significant in biology and nanomedicine research. Luminescent nanothermometry provides a noninvasive method of detecting the temperature in vivo with high sensitivity and high response speed. However, absorption and scattering in complex tissues limit the signal penetration depth and cause errors due to variation at different locations in vivo. In order to minimize these errors and monitor temperature in vivo, in the present work, we provided a strategy to fabricate a same-wavelength dual emission ratiometric upconversion luminescence nanothermometer based on a hybrid structure composed of upconversion emissive PbS quantum dots and Tm-doped upconversion nanoparticles. The ratiometric signal composed of two upconversion emissions working at the same wavelength, but different luminescent lifetimes, were decoded via a time-resolved technique. This nanothermometer improved the temperature monitoring ability and a thermal resolution and sensitivity of ~0.5 K and ~5.6% K−1 were obtained in vivo, respectively.


2001 ◽  
Vol 66 (10) ◽  
pp. 1473-1489 ◽  
Author(s):  
Drahomír Výprachtický ◽  
Věra Cimrová ◽  
Luďka Machová ◽  
Veronika Pokorná

Using the steady-state and time-resolved fluorescence spectroscopy, the behavior of "hairy-rod" oligo- and poly[2,5-bis(hexadecyloxy)-1,4-phenylene]s in tetrahydrofuran solutions was investigated. The materials were prepared by the Yamamoto coupling reaction using zinc as a reducing metal, the nickel(II)/triphenylphosphine complex as a catalyst, and 2,2'-bipyridine as a coligand. The appropriate oligomer fractions were separated by fractional precipitation and characterized by GPC and end group analysis. The fluorescence quantum yield of oligomers and polymers increased with their increasing conjugation length. The fluorescence emission spectra of polymers and longer oligomers exhibited one emission maximum at 390 nm with a single-exponential decay and fluorescence lifetimes (τ) around 1 ns. The substitution in positions 2 and 5 forces the adjacent backbone benzene units out of the plane, which results in twist angles 60-80°, and the bulky substituents exclude the cofacial sandwich-type configuration necessary for excimer formation. However, with shorter oligomers, another emission band at 460 nm appeared. Fluorescence decays at 460 nm were found to be double-exponential with longer excited-state lifetimes [e.g. τ1 = 6.9 ns (76%), τ2 = 2.4 ns (24%)]. With shorter oligomers (dimer, trimer), we assume a sandwich-type configuration with sufficiently close interchain distance and hence the excimer can form. Hydrophobic interactions of long aliphatic side chains in a polar medium play an important role in the excimer formation.


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