ArticleScientific reports2017
Applying phasor approach analysis of multiphoton FLIM measurements to probe the metabolic activity of three-dimensional in vitro cell culture models.
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 33 papers.
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Who cites it
33 citing papers in PubMed, 64 citations in OpenAlex.
- Optical diagnostics of liver tumors using an autofluorescence lifetime probe.Biomedical optics express · 2025Article
- Correlating NAD(P)H lifetime shifts to tamoxifen resistance in breast cancer cells: A metabolic screening study with time-resolved flow cytometry.Journal of innovative optical health sciences · 2025Article
- Phasor-FLIM and SHG imaging for quantitative analysis of lung cancer autofluorescence.Computational and structural biotechnology journal · 2025Article
- Label-free fluorescence lifetime imaging for the assessment of cell viability in living tumor fragments.Journal of biomedical optics · 2024Article
- Pioglitazone Phases and Metabolic Effects in Nanoparticle-Treated Cells Analyzed via Rapid Visualization of FLIM Images.Molecules (Basel, Switzerland) · 2024Article
- Advanced 3D imaging and organoid bioprinting for biomedical research and therapeutic applications.Advanced drug delivery reviews · 2024Review
- Probing organoid metabolism using fluorescence lifetime imaging microscopy (FLIM): The next frontier of drug discovery and disease understanding.Advanced drug delivery reviews · 2023Review
- Quantitative multiphoton imaging of cell metabolism, stromal fibers, and keratinization enables label-free discrimination of esophageal squamous cell carcinoma.Biomedical optics express · 2023Article
- Fluorescence Intensity and Fluorescence Lifetime Imaging Microscopies (FLIM) of Cell Differentiation in the Small Intestinal Organoids Using Cholera Toxin.Methods in molecular biology (Clifton, N.J.) · 2023Article
- Real-time open-source FLIM analysis.Frontiers in bioinformatics · 2023Article
- Single-cell imaging of α and β cell metabolic response to glucose in living human Langerhans islets.Communications biology · 2022Article
- Article
- Penetration Depth of Propylene Glycol, Sodium Fluorescein and Nile Red into the Skin Using Non-Invasive Two-Photon Excited FLIM.Pharmaceutics · 2022Article
- Accumulation and penetration behavior of hypericin in glioma tumor spheroids studied by fluorescence microscopy and confocal fluorescence lifetime imaging microscopy.Analytical and bioanalytical chemistry · 2022Article
- Toxicological Analysis of Hepatocytes Using FLIM Technique: In Vitro versus Ex Vivo Models.Cells · 2021Article
- Development and characterization of phasor-based analysis for FLIM to evaluate the metabolic and epigenetic impact of HER2 inhibition on squamous cell carcinoma cultures.Journal of biomedical optics · 2021Article
- New Extensibility and Scripting Tools in the ImageJ Ecosystem.Current protocols · 2021Article
- Multiphoton Microscopy and Mass Spectrometry for Revealing Metabolic Heterogeneity of HepatocytesSovremennye tekhnologii v meditsine · 2021Article
- Fluorescence Lifetime Imaging as a Noninvasive Tool to Study Plasmodium Falciparum Metabolism.Methods in molecular biology (Clifton, N.J.) · 2021Article
- In vivo non-invasive staining-free visualization of dermal mast cells in healthy, allergy and mastocytosis humans using two-photon fluorescence lifetime imaging.Scientific reports · 2020Article
Corrections and comments
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Authors and funding
5 authors at 5 institutions in 3 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Fluorescence lifetime imaging microscopy (FLIM) can measure and discriminate endogenous fluorophores present in biological samples. This study seeks to identify FLIM as a suitable method to non-invasively detect a shift in cellular metabolic activity towards glycolysis or oxidative phosphorylation in 3D Caco-2 models of colorectal carcinoma. These models were treated with potassium cyanide or hydrogen peroxide as controls, and epidermal growth factor (EGF) as a physiologically-relevant influencer of cell metabolic behaviour. Autofluorescence, attributed to nicotinamide adenine dinucleotide (NADH), was induced by two-photon laser excitation and its lifetime decay was analysed using a standard multi-exponential decay approach and also a novel custom-written code for phasor-based analysis. While both methods enabled detection of a statistically significant shift of metabolic activity towards glycolysis using potassium cyanide, and oxidative phosphorylation using hydrogen peroxide, employing the phasor approach required fewer initial assumptions to quantify the lifetimes of contributing fluorophores. 3D Caco-2 models treated with EGF had increased glucose consumption, production of lactate, and presence of ATP. FLIM analyses of these cultures revealed a significant shift in the contribution of protein-bound NADH towards free NADH, indicating increased glycolysis-mediated metabolic activity. This data demonstrate that FLIM is suitable to interpret metabolic changes in 3D in vitro models.
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