Chemical & Biomedical Imaging, Journal Year: 2025, Volume and Issue: unknown
Published: May 15, 2025
Language: Английский
Angewandte Chemie International Edition, Journal Year: 2023, Volume and Issue: 63(6)
Published: Nov. 13, 2023
Abstract Molecularly generated light, referred to here as “molecular light”, mainly includes bioluminescence, chemiluminescence, and Cerenkov luminescence. Molecular light possesses unique dual features of being both a molecule source light. Its molecular nature enables it be delivered molecules regions deep within the body, overcoming limitations natural sunlight physically sources like lasers LEDs. Simultaneously, its properties make valuable for applications such imaging, photodynamic therapy, photo‐oxidative photobiomodulation. In this review article, we provide an updated overview diverse discuss strengths weaknesses across various domains. Lastly, present forward‐looking perspectives on potential in realms photobiological mechanisms, therapeutic applications, While some these may considered bold contentious, our intent is inspire further innovations field applications.
Language: Английский
Citations
19
Annual Review of Analytical Chemistry, Journal Year: 2023, Volume and Issue: 16(1), P. 253 - 284
Published: June 14, 2023
Since the early 1990s, single-molecule detection in solution at room temperature has enabled direct observation of single biomolecules work real time and under physiological conditions, providing insights into complex biological systems that traditional ensemble methods cannot offer. In particular, recent advances tracking techniques allow researchers to follow individual their native environments for a timescale seconds minutes, revealing not only distinct pathways these take downstream signaling but also roles supporting life. this review, we discuss various imaging developed date, with an emphasis on advanced three-dimensional (3D) achieve ultrahigh spatiotemporal resolution provide sufficient working depths suitable molecules 3D tissue models. We then summarize observables can be extracted from trajectory data. Methods perform clustering analysis future directions are discussed.
Language: Английский
Citations
18
Journal of the American Chemical Society, Journal Year: 2024, Volume and Issue: 146(17), P. 11991 - 11999
Published: April 19, 2024
The complex dynamics and transience of assembly pathways in living systems complicate the understanding these molecular to nanoscale processes. Current technologies are unable track events leading onset assembly, where real-time information is imperative correlate their rich biology. Using a chemically designed pro-assembling molecule, we map its transformation into nanofibers fusion with endosomes form hollow fiber clusters. Tracked by phasor-fluorescence lifetime imaging (phasor-FLIM) epithelial cells (L929, A549, MDA-MB 231) correlative light-electron microscopy tomography (CLEM), spatiotemporal splicing shows time-correlated metabolic dysfunction. biological impact begins assembly-induced endosomal disruption that reduces glucose transport cells, which, turn, stymies mitochondrial respiration.
Language: Английский
Citations
7
Nature Communications, Journal Year: 2022, Volume and Issue: 13(1)
Published: Dec. 1, 2022
Abstract Fluorescence laser-scanning microscopy (LSM) is experiencing a revolution thanks to new single-photon (SP) array detectors, which give access an entirely set of information. Together with the blooming SP LSM techniques and development tailored there growing need for (i) DAQ systems capable handling high-throughput high-resolution photon information generated by these (ii) incorporating protocols in existing fluorescence LSMs. We developed open-source, low-cost, multi-channel time-tagging module (TTM) based on field-programmable gate that can tag parallel multiple events, 30 ps precision, synchronisation 4 ns precision. use TTM demonstrate live-cell super-resolved lifetime image scanning fluctuation spectroscopy. expect our BrightEyes-TTM will support community spreading SP-LSM many life science laboratories.
Language: Английский
Citations
23
Nature Methods, Journal Year: 2023, Volume and Issue: 20(2), P. 248 - 258
Published: Jan. 19, 2023
Abstract The expansion of fluorescence bioimaging toward more complex systems and geometries requires analytical tools capable spanning widely varying timescales length scales, cleanly separating multiple fluorescent labels distinguishing these from background autofluorescence. Here we meet challenging objectives for multispectral microscopy, combining hyperspectral phasors linear unmixing to create Hybrid Unmixing (HyU). HyU is efficient robust, quantitative signal separation even at low illumination levels. In dynamic imaging developing zebrafish embryos in mouse tissue, was able efficiently unmix labels, demanding volumetric timelapse settings. permits high range imaging, allowing simultaneous bright exogenous dim endogenous labels. This enables coincident studies tagged components, cellular behaviors metabolism within the same specimen, providing accurate insights into orchestrated complexity biological systems.
Language: Английский
Citations
15
Advanced Science, Journal Year: 2024, Volume and Issue: 11(32)
Published: June 20, 2024
Abstract Fluorescence lifetime imaging microscopy (FLIM) opens new dimensions for highly multiplexed in live cells and organisms using differences fluorescence to distinguish spectrally identical fluorescent probes. Here, a set of fluorescence‐activating absorption‐shifting tags (FASTs) capable modulating the embedded fluorogenic 4‐hydroxybenzylidene rhodanine (HBR) derivatives is described. It shown that changes FAST protein sequence can vary local environment chromophore lead significant lifetime. These lifetime‐modulating enable up three targets one spectral channel single HBR derivative zebrafish larvae. The combination multiplexing with allows successfully image six cells, opening great prospects multicolor multiplexing.
Language: Английский
Citations
6
Analytical Chemistry, Journal Year: 2022, Volume and Issue: 94(41), P. 14185 - 14194
Published: Oct. 3, 2022
The excited-state lifetime is an intrinsic property of fluorescent molecules that can be leveraged for multiplexed imaging. An advantage fluorescence lifetime-based multiplexing signals from multiple probes gathered simultaneously, whereas traditional spectral imaging typically requires images at different excitation and emission wavelengths. Additionally, spectra could both utilized to expand the capacity fluorescence. However, resolving exogenous molecular based exclusively on has been limited by technical challenges in analyzing data. phasor approach analysis offers a simple, graphical solution increasingly used assess endogenous cellular autofluorescence quantify metabolic factors. In this study, we employed FLIM quantitatively resolve three exogenous, antibody-targeted with similar properties information alone. First, demonstrated biomarkers were spatially restricted cell membrane, cytosol, or nucleus accurately distinguished using analysis. Next, successfully resolved quantified all targeted surface biomarkers. Finally, quantitation accuracy improved through intensity matching various probe-biomarker combinations, which will utility technique. Importantly, reconstructed each individual probe, as well overlay probes, single image. Our results demonstrate powerful tool simultaneous detection high sensitivity accuracy.
Language: Английский
Citations
22
Biomedicines, Journal Year: 2021, Volume and Issue: 9(11), P. 1528 - 1528
Published: Oct. 24, 2021
One of the challenges modern biology and medicine is to visualize biomolecules in their natural environment, real-time a non-invasive fashion, so as gain insight into physiological behavior highlight alterations pathological settings, which will enable devise appropriate therapeutic strategies. Genetically encoded fluorescent biosensors constitute class imaging agents that visualization biological processes events directly situ, preserving native context providing detailed localization dynamics cells. Real-time monitoring drug action specific cellular compartment, organ, or tissue type; ability screen at single-cell resolution; elimination false-positive results caused by low bioavailability not detected vitro testing methods are few obvious benefits using genetically screening. This review summarizes studies have been conducted last years toward fabrication for biomedical applications with comprehensive discussion on challenges, future trends, potential inputs needed improving them.
Language: Английский
Citations
27
Optics Letters, Journal Year: 2025, Volume and Issue: 50(5), P. 1484 - 1484
Published: Jan. 22, 2025
Label-free multiphoton microscopy is a powerful tool for investigating pristine biological specimens. This imaging modality leverages optical signals originating from the nonlinear response of native biomolecules to intense radiation, that allow localizing and quantifying constituents specimens, driving applications in biology medicine. However, since its inception over three decades ago, this approach has operated with narrowband detection scheme, relying on narrow bandwidths entire spectra derive contrast. scheme hinders analytical power microscope, preventing rigorous unmixing co-localized spectral overlap. In Letter, we shift paradigm broadband, demonstrating label-free hyperspectral We validate configuration by disentangling fresh murine tissues virtue spectra.
Language: Английский
Citations
0
Scientific Reports, Journal Year: 2025, Volume and Issue: 15(1)
Published: Feb. 14, 2025
Autofluorescence from endogenous biomolecules presents a significant challenge in immunofluorescence microscopy, often severely hindering the detection of specific fluorescence signals. While various techniques, including chemical- and quencher-based methods, photobleaching, digital-based protocols like autofluorescence image subtraction, have been developed to suppress autofluorescence, each approach has inherent limitations that restrict their efficacy across different tissue types. Fluorescence lifetime imaging microscopy (FLIM) offers powerful alternative by leveraging distinct lifetime-spectrum profiles fluorophores differentiate signals autofluorescence. However, traditional FLIM methods are slow hampered its use for suppression routine applications. In this study, we demonstrate GPU-accelerated high-speed can effectively separate samples, achieving high throughput meet demands biomedical clinical workflows. Furthermore, our findings show FLIM-based method enhances correlation images with immunohistochemistry data, outperforming other chemically-assisted photobleaching hyperspectral imaging. These results highlight potential speed improve reliability significantly.
Language: Английский
Citations
0