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Molecules
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Time-Resolved Luminescence Imaging and Spectroscopy
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Time-Resolved Luminescence Imaging and Spectroscopy

A special issue of Molecules (ISSN 1420-3049). This special issue belongs to the section "Photochemistry".

Deadline for manuscript submissions: closed (31 January 2022) | Viewed by 11974

Special Issue Editors

Dr. Riccardo Cicchi

E-Mail Website
Guest Editor
National Institute of Optics, National Research Council (CNR-INO), 1 Via Nello Carrara, I-50019 Sesto Fiorentino, Italy
Interests: biomedical optics; biophotonics; non-linear microscopy; fluorescence lifetime spectroscopy; tissue imaging
Special Issues, Collections and Topics in MDPI journals
Dr. Vladislav Shcheslavskiy

E-Mail
Co-Guest Editor
Institute of Experimental Oncology and Biomedical Technologies, Privolzhsky Research Medical University, 10/1, Minin and Pozharsky Sq., 603950 Nizhny Novgorod, Russia
Interests: time-correlated single photon counting, fluorescence lifetime imaging, non-linear optical microscopy and spectroscopy
Dr. João Lagarto

E-Mail Website
Assistant Guest Editor
Biophotonics Platform, Champalimaud Foundation, Lisbon, Portugal
Interests: biomedical optics; biophotonics; fluorescence; time-resolved spectroscopy; endoscopy; image-guided surgery; optical biopsy

Special Issue Information

Dear Colleagues,

Time-resolved luminescence methods are particularly attractive in various biological, biochemical, and biomedical applications, due to their sensitivity and specificity to biochemical, molecular, and physicochemical processes. Being dependent not only on the emitting fluorophore species but also on its molecular environment, time-resolved luminescence allows probing emitting molecules together with their molecular environment, providing both structural and functional information on the investigated specimen. The technological approach, generally based on time-gated or time-correlated single photon counting (TCSPC), has found application in tandem with various optical implementations, including confocal and multiphoton microscopy, macroscopic scanning, and wide-field imaging, and fiber-based spectroscopy and imaging. Time-resolved luminescence methods have been successfully applied to several research topics, ranging from molecular interaction and cell signaling to biological tissue characterization, frequently obtained through functional metabolic imaging of the NAD(P)H-FAD co-enzymes or through phosphorescence of oxygen molecular sensors. This Special Issue aims to review the developments of the technology, publish the latest advances, and illustrate the future prospective of the field.

Dr. Riccardo Cicchi
Guest Editor

Dr. Vladislav Shcheslavskiy
Co-Guest Editor

Dr. João Lagarto
Assistant Guest Editor

Manuscript Submission Information

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Keywords

  • Fluorescence and phosphorescence lifetime Imaging
  • Time-correlated single photon counting
  • Autofluorescence
  • Metabolism
  • FLIM-FRET

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Published Papers (4 papers)

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Research

11 pages, 2135 KiB  
Article
Automated Phasor Segmentation of Fluorescence Lifetime Imaging Data for Discriminating Pigments and Binders Used in Artworks
by Sara Mattana, Alice Dal Fovo, João Luís Lagarto, Maria Chiara Bossuto, Vladislav Shcheslavskiy, Raffaella Fontana and Riccardo Cicchi
Molecules 2022, 27(5), 1475; https://doi.org/10.3390/molecules27051475 - 22 Feb 2022
Cited by 7 | Viewed by 2299
Abstract
The non-invasive analysis of fluorescence from binders and pigments employed in mixtures in artworks is a major challenge in cultural heritage science due to the broad overlapping emission of different fluorescent species causing difficulties in the data interpretation. To improve the specificity of [...] Read more.
The non-invasive analysis of fluorescence from binders and pigments employed in mixtures in artworks is a major challenge in cultural heritage science due to the broad overlapping emission of different fluorescent species causing difficulties in the data interpretation. To improve the specificity of fluorescence measurements, we went beyond steady-state fluorescence measurements by resolving the fluorescence decay dynamics of the emitting species through time-resolved fluorescence imaging (TRFI). In particular, we acquired the fluorescence decay features of different pigments and binders using a portable and compact fibre-based imaging setup. Fluorescence time-resolved data were analysed using the phasor method followed by a Gaussian mixture model (GMM) to automatically identify the populations of fluorescent species within the fluorescence decay maps. Our results demonstrate that this approach allows distinguishing different binders when mixed with the same pigment as well as discriminating different pigments dispersed in a common binder. The results obtained could establish a framework for the analysis of a broader range of pigments and binders to be then extended to several other materials used in art production. The obtained results, together with the compactness and portability of the instrument, pave the way for future in situ applications of the technology on paintings. Full article
(This article belongs to the Special Issue Time-Resolved Luminescence Imaging and Spectroscopy)
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13 pages, 2724 KiB  
Article
Spectrally and Time-Resolved Fluorescence Imaging of 22-NBD-Cholesterol in Human Peripheral Blood Mononuclear Cells in Chronic Kidney Disease Patients
by Ingrid Lajdova, Livia Ovsonkova, Viera Spustova, Adrian Oksa, Dusan Chorvat, Anton Mateasik and Alzbeta Marcek Chorvatova
Molecules 2021, 26(22), 6800; https://doi.org/10.3390/molecules26226800 - 11 Nov 2021
Cited by 2 | Viewed by 2399
Abstract
The interaction of the fluorescent probe 22-NBD-cholesterol with membranes of human peripheral blood mononuclear cells (PBMC) was tested by time- and spectrally resolved fluorescence imaging to monitor the disturbance of lipid metabolism in chronic kidney disease (CKD) and its treatment with statins. Blood [...] Read more.
The interaction of the fluorescent probe 22-NBD-cholesterol with membranes of human peripheral blood mononuclear cells (PBMC) was tested by time- and spectrally resolved fluorescence imaging to monitor the disturbance of lipid metabolism in chronic kidney disease (CKD) and its treatment with statins. Blood samples from healthy volunteers (HV) and CKD patients, either treated or untreated with statins, were compared. Spectral imaging was done using confocal microscopy at 16 spectral channels in response to 458 nm excitation. Time-resolved imaging was achieved by time-correlated single photon counting (TCSPC) following excitation at 475 nm. The fluorescence of 22-NBD-cholesterol was mostly integrated into plasmatic membrane and/or intracellular membrane but was missing from the nuclear region. The presence of two distinct spectral forms of 22-NBD-cholesterol was uncovered, with significant variations between studied groups. In addition, two fluorescence lifetime components were unmasked, changing in CKD patients treated with statins. The gathered results indicate that 22-NBD-cholesterol may serve as a tool to study changes in the lipid metabolism of patients with CKD to monitor the effect of statin treatment. Full article
(This article belongs to the Special Issue Time-Resolved Luminescence Imaging and Spectroscopy)
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19 pages, 4131 KiB  
Article
Red Light-Emitting Water-Soluble Luminescent Iridium-Containing Polynorbornenes: Synthesis, Characterization and Oxygen Sensing Properties in Biological Tissues In Vivo
by Leonid N. Bochkarev, Yulia P. Parshina, Yana V. Gracheva, Tatyana A. Kovylina, Svetlana A. Lermontova, Larisa G. Klapshina, Aleksey N. Konev, Mikhail A. Lopatin, Maria M. Lukina, Anastasia D. Komarova, Vladislav I. Shcheslavskiy and Marina V. Shirmanova
Molecules 2021, 26(21), 6349; https://doi.org/10.3390/molecules26216349 - 20 Oct 2021
Cited by 6 | Viewed by 2713
Abstract
New water-soluble polynorbornenes P1–P4 containing oligoether, amino acid groups and luminophoric complexes of iridium(III) were synthesized by ring-opening metathesis polymerization. The polymeric products in organic solvents and in water demonstrate intense photoluminescence in the red spectral region. The polymers P1 and [...] Read more.
New water-soluble polynorbornenes P1–P4 containing oligoether, amino acid groups and luminophoric complexes of iridium(III) were synthesized by ring-opening metathesis polymerization. The polymeric products in organic solvents and in water demonstrate intense photoluminescence in the red spectral region. The polymers P1 and P3 with 1-phenylisoquinoline cyclometalating ligands in iridium fragments reveal 4–6 fold higher emission quantum yields in solutions than those of P2 and P4 that contain iridium complexes with 1-(thien-2-yl)isoquinoline cyclometalating ligands. The emission parameters of P1–P4 in degassed solutions essentially differ from those in the aerated solutions showing oxygen-dependent quenching of phosphorescence. Biological testing of P1 and P3 demonstrates that the polymers do not penetrate into live cultured cancer cells and normal skin fibroblasts and do not possess cytotoxicity within the concentrations and time ranges reasonable for biological studies. In vivo, the polymers display longer phosphorescence lifetimes in mouse tumors than in muscle, as measured using phosphorescence lifetime imaging (PLIM), which correlates with tumor hypoxia. Therefore, preliminary evaluation of the synthesized polymers shows their suitability for noninvasive in vivo assessments of oxygen levels in biological tissues. Full article
(This article belongs to the Special Issue Time-Resolved Luminescence Imaging and Spectroscopy)
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Graphical abstract

22 pages, 7047 KiB  
Article
Biocompatible Ir(III) Complexes as Oxygen Sensors for Phosphorescence Lifetime Imaging
by Ilya S. Kritchenkov, Anastasia I. Solomatina, Daria O. Kozina, Vitaly V. Porsev, Victor V. Sokolov, Marina V. Shirmanova, Maria M. Lukina, Anastasia D. Komarova, Vladislav I. Shcheslavskiy, Tatiana N. Belyaeva, Ilia K. Litvinov, Anna V. Salova, Elena S. Kornilova, Daniel V. Kachkin and Sergey P. Tunik
Molecules 2021, 26(10), 2898; https://doi.org/10.3390/molecules26102898 - 13 May 2021
Cited by 21 | Viewed by 3766
Abstract
Synthesis of biocompatible near infrared phosphorescent complexes and their application in bioimaging as triplet oxygen sensors in live systems are still challenging areas of organometallic chemistry. We have designed and synthetized four novel iridium [Ir(N^C)2(N^N)]+ complexes (N^C–benzothienyl-phenanthridine based cyclometalated ligand; [...] Read more.
Synthesis of biocompatible near infrared phosphorescent complexes and their application in bioimaging as triplet oxygen sensors in live systems are still challenging areas of organometallic chemistry. We have designed and synthetized four novel iridium [Ir(N^C)2(N^N)]+ complexes (N^C–benzothienyl-phenanthridine based cyclometalated ligand; N^N–pyridin-phenanthroimidazol diimine chelate), decorated with oligo(ethylene glycol) groups to impart these emitters’ solubility in aqueous media, biocompatibility, and to shield them from interaction with bio-environment. These substances were fully characterized using NMR spectroscopy and ESI mass-spectrometry. The complexes exhibited excitation close to the biological “window of transparency”, NIR emission at 730 nm, and quantum yields up to 12% in water. The compounds with higher degree of the chromophore shielding possess low toxicity, bleaching stability, absence of sensitivity to variations of pH, serum, and complex concentrations. The properties of these probes as oxygen sensors for biological systems have been studied by using phosphorescence lifetime imaging experiments in different cell cultures. The results showed essential lifetime response onto variations in oxygen concentration (2.0–2.3 μs under normoxia and 2.8–3.0 μs under hypoxia conditions) in complete agreement with the calibration curves obtained “in cuvette”. The data obtained indicate that these emitters can be used as semi-quantitative oxygen sensors in biological systems. Full article
(This article belongs to the Special Issue Time-Resolved Luminescence Imaging and Spectroscopy)
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