ArticleCytometry. Part A : the journal of the International Society for Analytical Cytology2003
Fluorescence lifetime-resolved pH imaging of living cells.
Article in Cytometry. Part A : the journal of the International Society for Analytical Cytology, 2003. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 66 papers.
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66 citing papers in PubMed, 189 citations in OpenAlex.
- Automated FLIM-FRET Segmentation Within RNP Condensates.Bio-protocol · 2026Article
- Luminescence lifetime imaging integrated into selective plane illumination microscopy for oxygen imaging in 3D cell cultures.Biomedical optics express · 2026Article
- Probing the photophysical properties of fluorescent proteins using photoacoustic pump-probe spectroscopy and imaging.Photoacoustics · 2025Article
- Review
- Fluorescence lifetime sorting reveals tunable enzyme interactions within cytoplasmic condensates.The Journal of cell biology · 2025Article
- Developing 1,4-Diethyl-1,2,3,4-tetrahydroquinoxalin-substituted Fluorogens Based on GFP Chromophore for Endoplasmic Reticulum and Lysosome Staining.International journal of molecular sciences · 2024Article
- Perinuclear damage from nuclear envelope deterioration elicits stress responses that contribute toScience advances · 2024Article
- Adaptation to sorbic acid in low sugar promotes resistance of yeast to the preservative.Heliyon · 2023Article
- Building Fluorescence Lifetime Maps Photon-by-Photon by Leveraging Spatial Correlations.ACS photonics · 2023Article
- Particle-based phasor-FLIM-FRET resolves protein-protein interactions inside single viral particles.Biophysical reports · 2023Article
- Direct Imaging of Local pH Reveals Bubble-Induced Mixing in a COACS sustainable chemistry & engineering · 2023Article
- Imaging the rotational mobility of carbon dot-gold nanoparticle conjugates using frequency domain wide-field time-resolved fluorescence anisotropy.Journal of biomedical optics · 2023Article
- Single-shot time-folded fluorescence lifetime imaging.Proceedings of the National Academy of Sciences of the United States of America · 2023Article
- Identifying lipid particle sub-types in liveFrontiers in chemistry · 2023Article
- Fluorescence lifetime image microscopy prediction with convolutional neural networks for cell detection and classification in tissues.PNAS nexus · 2022Article
- Design and characterization of a time-domain optical tomography platform for mesoscopic lifetime imaging.Biomedical optics express · 2022Article
- Penetration Depth of Propylene Glycol, Sodium Fluorescein and Nile Red into the Skin Using Non-Invasive Two-Photon Excited FLIM.Pharmaceutics · 2022Article
- Article
- Fluorescence lifetime-based pH mapping of tumors in vivo using genetically encoded sensor SypHerRed.Biophysical journal · 2022Article
- Monte Carlo-based data generation for efficient deep learning reconstruction of macroscopic diffuse optical tomography and topography applications.Journal of biomedical optics · 2022Article
6 more citing papers are in PubMed but not listed here.
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Authors and funding
3 authors at 1 institution in 1 country.
Funding
Abstract
backgroundThe regulation and maintenance of intracellular pH are critical to diverse metabolic functions of the living cells. Fluorescence time-resolved techniques and instrumentations have advanced rapidly and enabled the imaging of intracellular pH based on the fluorescence lifetimes.
methodsThe frequency-domain fluorescence lifetime imaging microscopy (FLIM) and fluorophores displaying appropriate pH-dependent lifetime sensitivities were used to determine the temporal and spatial pH distributions in the cytosol and vesicular compartment lysosomes.
resultsWe found that cytosolic pH levels are different in 3T3 fibroblasts, Chinese hamster ovary (CHO) cells, and MCF-7 cells when using the pH probe carboxy-SNAFL2. We also tracked the transient cytosolic pH changes in the living CHO cells after treatments with proton pump inhibitors, ion exchanger inhibitors, and weak base and acid. The intracellular lysosomal pH was determined with the acidic lifetime probes DM-NERF dextrans, OG-514 carboxylic acid dextrans, and LysoSensor DND-160. Our results showed that the resting lysosomal pH value obtained from the 3T3 fibroblasts was between 4.5 and 4.9. The increase of lysosomal pH induced by the treatments with proton pump inhibitor and ionophores also were observed in our FLIM measurements.
conclusionsOur lifetime-based pH imaging data suggested that FLIM can measure the intracellular pH of the resting cells and follow the pH fluctuations inside the cells after environmental perturbations. To improve the z-axis resolution to the intracellular lifetime-resolved images, we are investigating the implementation of the pseudo-confocal capability to our current FLIM apparatus.
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