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
Plant Sciences
Elena Kristin Petutschnig, Leon Pierdzig, Josephine Mittendorf, Jule Meret Niebisch, Volker Lipka
Summary: Elucidating protein-protein interactions is crucial for understanding molecular processes. A novel combination of fluorescence proteins, mCitrine and mScarlet-I, is suitable for FLIM-FRET studies in stably transformed plants, allowing detection of low abundance proteins and facilitating co-localization and co-immunoprecipitation experiments.
JOURNAL OF EXPERIMENTAL BOTANY
(2023)
Article
Chemistry, Applied
Laura Espinar-Barranco, Jose Manuel Paredes, Angel Orte, Luis Crovetto, Emilio Garcia-Fernandez
Summary: In this study, a sensor with solvatofluorochromic property was used to investigate the initial stages of beta-amyloid peptide aggregation in Alzheimer's disease. By detecting changes in environmental polarity, the hydrophobicity of the aggregates can be quantified and different types of aggregates can be distinguished using advanced fluorescence spectroscopy and imaging techniques.
Review
Biochemistry & Molecular Biology
Dorothy Koveal
Summary: Genetically encoded fluorescent biosensors are used to measure chemical changes in single cells rapidly. These biosensors are mainly used in tracking neural activity and neurotransmitter release, but their use and data interpretation for studying brain metabolism pose challenges. Many biosensors are prone to interferences that can lead to ambiguous results. This review discusses current methods of biosensor quantitation, focusing on cellular interferences, methods to avoid false inferences, and recent advances in sensor optimization.
JOURNAL OF NEUROCHEMISTRY
(2023)
Article
Biochemistry & Molecular Biology
Tatiana R. Simonyan, Elena A. Protasova, Anastasia Mamontova, Aleksander M. Shakhov, Konstantin A. Lukyanov, Eugene G. Maksimov, Alexey M. Bogdanov
Summary: The study demonstrates that a green-fluorescence-protein-based emitter has high sensitivity in alkaline pH range, enabling accurate measurement of mitochondrial pH in live cells.
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
(2022)
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
Polymer Science
Koushik Bhattacharya, Moumita Kundu, Subhayan Das, Sarthik Samanta, Sib Sankar Roy, Mahitosh Mandal, Nikhil K. Singha
Summary: A functional fluorescent sensor has been developed in this study, which can distinguish cancer cells from normal cells by monitoring the pH change.
MACROMOLECULAR RAPID COMMUNICATIONS
(2023)
Article
Chemistry, Multidisciplinary
Jorrit Bleeker, Aron P. Kahn, Lorenz M. Baumgartner, Ferdinand C. Grozema, David A. Vermaas, Wolter F. Jager
Summary: Spatiotemporal pH imaging using fluorescence lifetime imaging microscopy (FLIM) is an excellent technique for investigating dynamic (electro)chemical processes. However, current probes for FLIM are not suitable for high pH values. In this study, we developed dedicated pH probes based on 1-methyl-7-amino-quinolinium fluorophore, which possesses high fluorescence lifetime and quantum yield, (photo)stability, and water solubility. The flexible fluorophore-spacer-receptor architecture allows tunable probe lifetimes in the pH range of 5.5-11, and deprotonation of the aromatic amine at the quinolinium core enables additional fluorescence lifetime response at pH values of 11-13.
Article
Multidisciplinary Sciences
Brian Tenner, Jason Z. Zhang, Yonghoon Kwon, Veronica Pessino, Siyu Feng, Bo Huang, Sohum Mehta, Jin Zhang
Summary: FluoSTEPs is a novel class of biosensors that combines self-complementing split green fluorescent protein, CRISPR-mediated knock-in, and fluorescence resonance energy transfer technology to probe compartmentalized signaling dynamics in situ. It can simultaneously highlight endogenous microdomains and report domain-specific, real-time signaling events, including kinase activities, guanosine triphosphatase activation, and second messenger dynamics. FluoSTEPs have been shown to be useful in probing spatiotemporal regulation within endogenous signaling architectures, as demonstrated by a FluoSTEP for 3',5'-cyclic adenosine monophosphate (cAMP) revealing distinct cAMP dynamics within clathrin microdomains in response to stimulation of G protein-coupled receptors.
Article
Spectroscopy
Luo Lin-lin, Niu Jing-jing, Mo Bei-xin, Lin Dan-ying, Liu Lin
Summary: Cell is the basic unit of structure and biological activities, and cellular processes involve spatiotemporally interactions of biochemical components. Traditional biochemical methods face challenges in evaluating molecular interactions in living cells, while the advancement in optical technology, such as FRET-FLIM, offers new genetic research tools for real-time exploration. FRET-FLIM technology provides unique advantages in detecting and analyzing dynamic interactions of biomolecules in living cells, achieving high resolution visualization, sensitivity, and fast analysis.
SPECTROSCOPY AND SPECTRAL ANALYSIS
(2021)
Review
Plant Sciences
Zhikun Duan, Kaiwen Li, Wenwen Duan, Junli Zhang, Jingjing Xing
Summary: This article highlights the importance of using FRET technology to study interactions of plant membrane proteins, providing an overview of its applications in quantifying dynamic interactions and assemblies, as well as sensors for quantifying signaling molecule homeostasis and kinase activity. The recent applications of advanced FRET sensors in probing membrane protein interactions, stoichiometry, and clustering have shed light on the complex biological functions of membrane proteins in living plant cells.
JOURNAL OF EXPERIMENTAL BOTANY
(2022)
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
Chemistry, Multidisciplinary
Yasmina Bousmah, Hana Valenta, Giulia Bertolin, Utkarsh Singh, Valerie Nicolas, Helene Pasquier, Marc Tramier, Fabienne Merola, Marie Erard
Summary: The yellow fluorescent protein tdLanYFP, derived from a tetrameric protein from cephalochordate Branchiostoma lanceolatum, exhibits excellent quantum yield, extinction coefficient, and photostability, making it a bright dimeric fluorescent protein with compatibility for imaging cellular structures at subdiffraction resolution. Its low pK(1/2) value of 3.9 also allows for excellent performance at acidic pH levels, making it a valuable tag for FRET experiments in various biosensing modalities.
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)
Review
Chemistry, Analytical
Namdoo Kim, Seunghan Shin, Se Won Bae
Summary: cAMP serves a crucial role in signal transduction pathways, with research focusing on its dynamics leading to insights for drug development and disease treatment. To enable real-time non-invasive imaging, genetically-encoded sensors based on fluorescent proteins and luciferases have been developed as powerful tools.
