ArticlePloS one2013
Parallelized TCSPC for dynamic intravital fluorescence lifetime imaging: quantifying neuronal dysfunction in neuroinflammation.
Article in PloS one, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 34 papers.
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Who cites it
34 citing papers in PubMed, 69 citations in OpenAlex.
- Tracking CNS and systemic sources of oxidative stress during the course of chronic neuroinflammation.Acta neuropathologica · 2015Trial
- Monitoring Calcium Dynamics in Pig Intestinal Organoids with FLIM and FRET.Methods in molecular biology (Clifton, N.J.) · 2027Article
- Multimodal Longitudinal Optical Imaging Reveals Optic Neuritis Preceding Retinal Pathology in Experimental Autoimmune Encephalomyelitis.Neurology(R) neuroimmunology & neuroinflammation · 2025Article
- Probing organoid metabolism using fluorescence lifetime imaging microscopy (FLIM): The next frontier of drug discovery and disease understanding.Advanced drug delivery reviews · 2023Review
- Combined FRET-FLIM and NAD(P)H FLIM to Analyze B Cell Receptor Signaling Induced Metabolic Activity of Germinal Center B Cells In Vivo.Methods in molecular biology (Clifton, N.J.) · 2023Article
- Two-Photon Excitation Spectra of Various Fluorescent Proteins within a Broad Excitation Range.International journal of molecular sciences · 2022Article
- Instant FLIM enables 4DOptica · 2021Article
- Luminescence lifetime imaging of three-dimensional biological objects.Journal of cell science · 2021Article
- Fast Gating for Raman Spectroscopy.Sensors (Basel, Switzerland) · 2021Review
- Article
- A Guide to Fluorescence Lifetime Microscopy and Förster's Resonance Energy Transfer in Neuroscience.Current protocols in neuroscience · 2020Review
- Fluorescence lifetime imaging microscopy: fundamentals and advances in instrumentation, analysis, and applications.Journal of biomedical optics · 2020Review
- Evaluation of cell metabolic adaptation in wound and tumour by Fluorescence Lifetime Imaging Microscopy.Scientific reports · 2020Article
- High-throughput, multi-parametric, and correlative fluorescence lifetime imaging.Methods and applications in fluorescence · 2020Review
- Teriflunomide Does Not Change Dynamics of Nadph Oxidase Activation and Neuronal Dysfunction During Neuroinflammation.Frontiers in molecular biosciences · 2020Article
- Machine Learning Methods for Fluorescence Lifetime Imaging (FLIM) Based Label-Free Detection of Microglia.Frontiers in neuroscience · 2020Article
- Systematic Enzyme Mapping of Cellular Metabolism by Phasor-Analyzed Label-Free NAD(P)H Fluorescence Lifetime Imaging.International journal of molecular sciences · 2019Article
- Wide-field time-correlated single photon counting-based fluorescence lifetime imaging microscopy.Nuclear instruments & methods in physics research. Section A, Accelerators, spectrometers, detectors and associated equipment · 2019Review
- Fast single-cell biochemistry: theory, open source microscopy and applications.Methods and applications in fluorescence · 2019Article
- Recent trends in two-photon auto-fluorescence lifetime imaging (2P-FLIM) and its biomedical applications.Biomedical engineering letters · 2019Review
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Authors and funding
14 authors at 6 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Two-photon laser-scanning microscopy has revolutionized our view on vital processes by revealing motility and interaction patterns of various cell subsets in hardly accessible organs (e.g. brain) in living animals. However, current technology is still insufficient to elucidate the mechanisms of organ dysfunction as a prerequisite for developing new therapeutic strategies, since it renders only sparse information about the molecular basis of cellular response within tissues in health and disease. In the context of imaging, Förster resonant energy transfer (FRET) is one of the most adequate tools to probe molecular mechanisms of cell function. As a calibration-free technique, fluorescence lifetime imaging (FLIM) is superior for quantifying FRET in vivo. Currently, its main limitation is the acquisition speed in the context of deep-tissue 3D and 4D imaging. Here we present a parallelized time-correlated single-photon counting point detector (p-TCSPC) (i) for dynamic single-beam scanning FLIM of large 3D areas on the range of hundreds of milliseconds relevant in the context of immune-induced pathologies as well as (ii) for ultrafast 2D FLIM in the range of tens of milliseconds, a scale relevant for cell physiology. We demonstrate its power in dynamic deep-tissue intravital imaging, as compared to multi-beam scanning time-gated FLIM suitable for fast data acquisition and compared to highly sensitive single-channel TCSPC adequate to detect low fluorescence signals. Using p-TCSPC, 256×256 pixel FLIM maps (300×300 µm(2)) are acquired within 468 ms while 131×131 pixel FLIM maps (75×75 µm(2)) can be acquired every 82 ms in 115 µm depth in the spinal cord of CerTN L15 mice. The CerTN L15 mice express a FRET-based Ca-biosensor in certain neuronal subsets. Our new technology allows us to perform time-lapse 3D intravital FLIM (4D FLIM) in the brain stem of CerTN L15 mice affected by experimental autoimmune encephalomyelitis and, thereby, to truly quantify neuronal dysfunction in neuroinflammation.
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Registered trials
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.