Chemical & Biomedical Imaging, Journal Year: 2025, Volume and Issue: unknown
Published: May 15, 2025
Language: Английский
Nature Methods, Journal Year: 2021, Volume and Issue: 19(1), P. 65 - 70
Published: Dec. 16, 2021
Abstract Self-labeling protein tags such as HaloTag are powerful tools that can label fusion proteins with synthetic fluorophores for use in fluorescence microscopy. Here we introduce variants either increased or decreased brightness and lifetime compared HaloTag7 when labeled rhodamines. Combining these enabled live-cell multiplexing of three cellular targets one spectral channel using a single fluorophore the generation lifetime-based biosensor. Additionally, brightest variant showed up to 40% higher imaging applications.
Language: Английский
Citations
124
Nature Reviews Molecular Cell Biology, Journal Year: 2024, Volume and Issue: 25(6), P. 443 - 463
Published: Feb. 20, 2024
Language: Английский
Citations
26
Nature Communications, Journal Year: 2022, Volume and Issue: 13(1)
Published: May 5, 2022
Abstract Ultra-multiplexed fluorescence imaging requires the use of spectrally overlapping fluorophores to label proteins and then unmix images fluorophores. However, doing this remains a challenge, especially in highly heterogeneous specimens, such as brain, owing high degree variation emission spectra specimens. Here, we propose PICASSO, which enables more than 15-color spatially single round without using any reference spectra. PICASSO an equal number fluorophores, advanced multiplexed imaging, even with bandpass filter-based microscopy. We show that can be used achieve strong multiplexing capability diverse applications. By combining cyclic immunofluorescence staining, 45-color mouse brain three cycles. provides tool for accessibility accuracy broad range researchers.
Language: Английский
Citations
65
Nature Communications, Journal Year: 2022, Volume and Issue: 13(1)
Published: Jan. 10, 2022
Abstract Multiplexed mRNA profiling in the spatial context provides new information enabling basic research and clinical applications. Unfortunately, existing transcriptomics methods are limited due to either low multiplexing or complexity. Here, we introduce a spatialomics technology, termed Multi Omic Single-scan Assay with Integrated Combinatorial Analysis (MOSAICA), that integrates situ labeling of protein markers cells tissues combinatorial fluorescence spectral lifetime encoded probes, time-resolved imaging, machine learning-based decoding. We demonstrate MOSAICA’s scalability detecting 10-plex targets fixed colorectal cancer using five fluorophores facile error-detection removal autofluorescence. analysis is strongly correlated sequencing data (Pearson’s r = 0.96) was further benchmarked RNAscope TM LGC Stellaris . apply MOSAICA for multiplexed melanoma Formalin-Fixed Paraffin-Embedded (FFPE) tissues. finally simultaneous co-detection cells.
Language: Английский
Citations
54
Communications Biology, Journal Year: 2024, Volume and Issue: 7(1)
Published: March 26, 2024
Abstract For generations researchers have been observing the dynamic processes of life through lens a microscope. This has offered tremendous insights into biological phenomena that span multiple orders time- and length-scales ranging from pure magic molecular reorganization at membrane immune cells, to cell migration differentiation during development or wound healing. Standard fluorescence microscopy techniques offer glimpses such in vitro, however, when applied intact systems, they are challenged by reduced signal strengths signal-to-noise ratios result deeper imaging. As remedy, two-photon excitation (TPE) takes special place, because it allows us investigate vivo, their natural environment, even living animal. Here, we review fundamental principles underlying TPE aimed basic advanced users interested adopting for intravital We focus on applications neurobiology, present current trends towards faster, wider imaging, discuss combination with photon counting technologies metabolic imaging spectroscopy, as well highlight outstanding issues drawbacks application these methodologies.
Language: Английский
Citations
16
Reviews of Modern Physics, Journal Year: 2024, Volume and Issue: 96(2)
Published: June 5, 2024
For centuries, human fascination with the living world motivated development of tools for visualizing life's events at spatiotemporal scales beyond our visual range. While all optical microscopes use light to probe object interest, fluorescence can discern between and background molecular scale. At this scale, stochastic properties are fundamental interpreting microscopy data. Accordingly quantitative methods that enable such interpretation necessitate perspective statistical concepts. The physical-optical principles governing formation fluorescent images modeling these while accounting stochasticity measurements reviewed.
Language: Английский
Citations
13
Nature Reviews Methods Primers, Journal Year: 2024, Volume and Issue: 4(1)
Published: Nov. 7, 2024
Language: Английский
Citations
9
Advanced Drug Delivery Reviews, Journal Year: 2025, Volume and Issue: 218, P. 115521 - 115521
Published: Jan. 22, 2025
Language: Английский
Citations
1
ACS Chemical Biology, Journal Year: 2022, Volume and Issue: 17(6), P. 1321 - 1327
Published: May 18, 2022
Fluorescence lifetime multiplexing requires fluorescent probes with distinct fluorescence lifetimes but similar spectral properties. Even though synthetic for many cellular targets are available multicolor live-cell microscopy, few of them have been characterized their use in multiplexing. Here, we demonstrate that, from a panel 18 probes, eight pairwise combinations suitable living mammalian cell lines. Moreover, combining multiple pairs different channels enables us to image four and the help self-labeling protein tags up biological targets, effectively doubling number observable targets. The combination is thus powerful approach imaging.
Language: Английский
Citations
32
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