Review
Chemistry, Analytical
Cong Quang Vu, Satoshi Arai
Summary: Genetically encoded fluorescence lifetime biosensors provide a powerful tool for quantitative imaging, enabling precise measurement of cellular metabolites, molecular interactions, and dynamic cellular processes. This review gives an overview of the principles, applications, and advancements in quantitative imaging with genetically encoded fluorescence lifetime biosensors using fluorescence lifetime imaging microscopy (go-FLIM), highlighting the distinct advantages of fluorescence lifetime-based measurements.
Article
Engineering, Environmental
Adrian Monteleone, Folker Wenzel, Heinz Langhals, Daniel Dietrich
Summary: The FLIM method was tested for identification and characterization of six types of plastics, showing significant differentiation in 94.55% of the comparisons. Results suggest that FLIM has the potential for sub-micrometer plastic characterization and allows for visual 3D-sectioning of samples, which could be important for identifying and characterizing plastics in tissue and environmental samples.
JOURNAL OF ENVIRONMENTAL CHEMICAL ENGINEERING
(2021)
Article
Biochemistry & Molecular Biology
Andrew H. A. Clayton
Summary: The dynamics of condensed matter can be studied using the time-dependent Stokes shift of a fluorescent probe. The time-dependent spectral correlation function is described by spectral relaxation correlation times that characterize the relaxation processes around the excited-state probe. Phasor plot is a powerful tool to analyze time and frequency-domain data, providing spatial maps of spectral dynamics in complex structures like living cells. The analysis workflow also addresses the extraction of initial and relaxed generalized polarization and improved methods for model discrimination and parameter extraction in complex spectral relaxation phenomena.
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
(2023)
Article
Obstetrics & Gynecology
Jaimin S. Shah, Marta Venturas, Tim H. Sanchez, Alan S. Penzias, Daniel J. Needleman, Denny Sakkas
Summary: The study found significant metabolic differences between euploid and aneuploid human blastocysts using FLIM, indicating potential for FLIM as a non-invasive clinical tool to assist in identifying ploidy status.
HUMAN REPRODUCTION
(2022)
Article
Biochemistry & Molecular Biology
Polina Vishnyakova, Elena Nikonova, Enar Jumaniyazova, Ilya Solovyev, Anastasia Kirillova, Maria Farmakovskaya, Alexander Savitsky, Evgeny Shirshin, Gennady Sukhikh, Timur Fatkhudinov
Summary: In this study, the metabolic status of spermatozoa from healthy donors was assessed using fluorescence lifetime imaging microscopy (FLIM). It was found that FLIM could be used to separate and group male germ cells based on their metabolic activity, which corresponded with the sperm motility measured in standard spermogram tests.
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
(2023)
Article
Chemistry, Multidisciplinary
Martin Priessner, Peter A. Summers, Benjamin W. Lewis, Magdalena Sastre, Liming Ying, Marina K. Kuimova, Ramon Vilar
Summary: A new optical probe based on BODIPY has been developed, which exhibits fluorescence intensity switch-on upon binding to copper(I) and can be used to visualize copper(I) pools in lysosomes of live cells.
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
(2021)
Article
Chemistry, Physical
Cecilia Zaza, German Chiarelli, Ludovit P. Zweifel, Mauricio Pilo-Pais, Evangelos Sisamakis, Fabio Barachati, Fernando D. Stefani, Guillermo P. Acuna
Summary: Fluorescence Resonance Energy Transfer (FRET)-based approaches are unique tools for sensing the immediate surroundings and interactions of (bio)molecules. FRET imaging and Fluorescence Lifetime Imaging Microscopy (FLIM) enable the visualization of the spatial distribution of molecular interactions and functional states. However, conventional FLIM and FRET imaging provide average information over an ensemble of molecules within a diffraction-limited volume, which limits the spatial information, accuracy, and dynamic range of the observed signals. Here, an approach to obtain super-resolved FRET imaging based on single-molecule localization microscopy using an early prototype of a commercial time-resolved confocal microscope is demonstrated. DNA Points Accumulation for Imaging in Nanoscale Topography with fluorogenic probes provides a suitable combination of background reduction and binding kinetics compatible with the scanning speed of usual confocal microscopes. A single laser is used to excite the donor, a broad detection band is employed to retrieve both donor and acceptor emission, and FRET events are detected from lifetime information.
Article
Biochemistry & Molecular Biology
Adrian Monteleone, Weronika Schary, Folker Wenzel, Heinz Langhals, Daniel R. Dietrich
Summary: The study introduced a new method FLIM and phasor analysis for fast, label-free, and sensitive identification and differentiation of plastics, which represents a significant advancement in addressing the issue of plastic pollution.
CHEMICO-BIOLOGICAL INTERACTIONS
(2021)
Article
Nanoscience & Nanotechnology
Subhabrata Ghosh, Jennifer A. Hollingsworth, Jose Ignacio Gallea, Somak Majumder, Joerg Enderlein, Alexey Chizhik
Summary: This article reports on the proof of principle measurements of a super-resolution imaging method based on the excitation field density-dependent lifetime modulation of semiconductor nanocrystals. Experimental results demonstrate that the method can achieve a spatial resolution of several tens of nanometers at moderate fluorescence excitation intensity.
Article
Plant Sciences
Nina Gloeckner, Sven zur Oven-Krockhaus, Leander Rohr, Frank Wackenhut, Moritz Burmeister, Friederike Wanke, Eleonore Holzwart, Alfred J. Meixner, Sebastian Wolf, Klaus Harter
Summary: Protein-protein interaction studies are crucial for understanding cellular signaling. This study used three-fluorophore FRET and FRET-fluorescence lifetime imaging microscopy techniques to demonstrate the formation of a ternary complex involving Receptor-Like Protein 44, Brassinosteroid Insensitive 1, and Brassinosteroid Insensitive 1 Associated Kinase 1 in living plant cells. The researchers also found that this complex is localized in a distinct plasma membrane nanodomain.
Article
Nanoscience & Nanotechnology
Janet E. Sorrells, Rishyashring R. Iyer, Lingxiao Yang, Elisabeth M. Martin, Geng Wang, Haohua Tu, Marina Marjanovic, Stephen A. Boppart
Summary: This paper presents a computational photon counting method using a hybrid photodetector and multithreshold peak detection, which enables faster imaging. By counting instances where photons arrive at the detector within the detector response time, the method accurately characterizes the intensity and fluorescence lifetime of samples.
Article
Chemistry, Multidisciplinary
Linda Sistemich, Phillip Galonska, Jan Stegemann, Julia Ackermann, Sebastian Kruss
Summary: Single-walled carbon nanotubes (SWCNTs) are versatile building blocks for biosensors that can respond to analytes by a change in fluorescence. However, intensity-based signals are easily affected by external factors. In this study, we demonstrate the use of fluorescence lifetime imaging microscopy (FLIM) to overcome this issue and utilize SWCNT-based sensors for detecting dopamine.
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
(2023)
Article
Nanoscience & Nanotechnology
Xavier Michalet
Summary: This study revisited the time-resolved analysis of periodically excited luminescence decays by the phasor method in the presence of time-gating or binning. Analytical expressions for discrete configurations of square gates were derived, and the effects of instrument response function offset, decay truncation, and gate shape were discussed. Modified expressions for the phase and modulus lifetimes were provided for some simple cases, along with a discussion of a modified phasor calibration approach.
Article
Genetics & Heredity
Jieqiong Lou, Ashleigh Solano, Zhen Liang, Elizabeth Hinde
Summary: The study introduces a phasor approach to fluorescence lifetime imaging microscopy (FLIM) that can directly measure chromatin network architecture and detect changes in this structural framework upon induction of DNA double-strand breaks (DSBs) within an intact nucleus. By combining FRET with immunofluorescence, this technology allows for exploration of any heterogeneity that exists in chromatin structure at spatially distinct and genetically induced DSBs.
FRONTIERS IN GENETICS
(2021)
Article
Biochemistry & Molecular Biology
Thomas Kellerer, Janko Janusch, Christian Freymueller, Adrian Ruehm, Ronald Sroka, Thomas Hellerer
Summary: This study utilized two different fluorescence lifetime measurement techniques to investigate the factors affecting fluorescence lifetime results, showing consistent results and revealing the strongest influencing factor on fluorescence lifetime. It also provides quantitative guidance on how to choose the most suitable technique and perform experiments correctly to obtain consistent fluorescence lifetime results.
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
(2022)
Article
Oncology
Yuxin Liu, Swati Bhardwaj, Keith Sigel, John Winters, Joseph Terlizzi, Michael M. Gaisa
Summary: This study investigated the prevalence and severity of anal HPV disease among MSM LWH under the age of 35, finding a high prevalence of HPV infection and precancer but no cases of invasive anal cancer. This supports the adoption of age-based anal cancer screening for this population.
INTERNATIONAL JOURNAL OF CANCER
(2024)
Correction
Oncology
J. Gu, S. Xie, S. Wang
INTERNATIONAL JOURNAL OF CANCER
(2024)