ArticleScientific reports2017
Synergistic Strategy for Multicolor Two-photon Microscopy: Application to the Analysis of Germinal Center Reactions In Vivo.
Article in Scientific reports, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 31 papers.
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Who cites it
31 citing papers in PubMed, 60 citations in OpenAlex.
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- Supercontinuum-tailoring multicolor imaging reveals spatiotemporal dynamics of heterogeneous tumor evolution.Nature communications · 2024Article
- Optimized intravital three-photon imaging of intact mouse tibia links plasma cell motility to functional states.iScience · 2024Article
- BINGO: a blind unmixing algorithm for ultra-multiplexing fluorescence images.Bioinformatics (Oxford, England) · 2024Article
- Silica-Encapsulated Perovskite Nanocrystals for X-ray-Activated Singlet Oxygen Production and Radiotherapy Application.ACS energy letters · 2023Article
- HyU: Hybrid Unmixing for longitudinal in vivo imaging of low signal-to-noise fluorescence.Nature methods · 2023Article
- SUFI: an automated approach to spectral unmixing of fluorescent multiplex images captured in mouse and post-mortem human brain tissues.BMC neuroscience · 2023Article
- Two-Photon Excitation Spectra of Various Fluorescent Proteins within a Broad Excitation Range.International journal of molecular sciences · 2022Article
- Super-multiplexing excitation spectral microscopy with multiple fluorescence bands.Biomedical optics express · 2022Article
- [The DRFZ-a pioneer in research on the interaction between immune and stromal cells during de- and regeneration of the musculoskeletal system].Zeitschrift fur Rheumatologie · 2022Review
- Recycling of memory B cells between germinal center and lymph node subcapsular sinus supports affinity maturation to antigenic drift.Nature communications · 2022Article
- NAD(P)H fluorescence lifetime imaging of live intestinal nematodes reveals metabolic crosstalk between parasite and host.Scientific reports · 2022Article
- Diamond Raman laser and Yb fiber amplifier forBiomedical optics express · 2022Article
- Two-Photon Absorption: An Open Door to the NIR-II Biological Window?Frontiers in chemistry · 2022Review
- Review
- Single-laser-based simultaneous four-wavelength excitation source for femtosecond two-photon fluorescence microscopy.Biomedical optics express · 2021Article
Corrections and comments
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Authors and funding
13 authors at 3 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Simultaneous detection of multiple cellular and molecular players in their native environment, one of the keys to a full understanding of immune processes, remains challenging for in vivo microscopy. Here, we present a synergistic strategy for spectrally multiplexed in vivo imaging composed of (i) triple two-photon excitation using spatiotemporal synchronization of two femtosecond lasers, (ii) a broad set of fluorophores with emission ranging from blue to near infrared, (iii) an effective spectral unmixing algorithm. Using our approach, we simultaneously excite and detect seven fluorophores expressed in distinct cellular and tissue compartments, plus second harmonics generation from collagen fibers in lymph nodes. This enables us to visualize the dynamic interplay of all the central cellular players during germinal center reactions. While current in vivo imaging typically enables recording the dynamics of 4 tissue components at a time, our strategy allows a more comprehensive analysis of cellular dynamics involving 8 single-labeled compartments. It enables to investigate the orchestration of multiple cellular subsets determining tissue function, thus, opening the way for a mechanistic understanding of complex pathophysiologic processes in vivo. In the future, the design of transgenic mice combining a larger spectrum of fluorescent proteins will reveal the full potential of our method.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.