ArticleBiophysical journal2008
Refractive index sensing of green fluorescent proteins in living cells using fluorescence lifetime imaging microscopy.
Article in Biophysical journal, 2008. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 42 papers.
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42 citing papers in PubMed, 134 citations in OpenAlex.
- Meta-surface plasmon resonance microscopy for sensitive, high-throughput, and long-term label-free analysis of cell adhesion dynamics.Journal of nanobiotechnology · 2025Article
- Evaluation of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) using a high figure-of-merit plasmonic multimode refractive index optical sensor.Scientific reports · 2024Article
- Fluorescent protein lifetimes report densities and phases of nuclear condensates during embryonic stem-cell differentiation.Nature communications · 2023Article
- Label-Free Imaging Techniques to Evaluate Metabolic Changes Caused by Toxic Liver Injury in PCLS.International journal of molecular sciences · 2023Article
- FLIM imaging revealed spontaneous osteogenic differentiation of stem cells on gradient pore size tissue-engineered constructs.Stem cell research & therapy · 2023Article
- Irreversible alteration of extracellular vesicle and cell-free messenger RNA profiles in human plasma associated with blood processing and storage.Scientific reports · 2022Article
- Sensitive Detection of SARS-CoV-2 Using a Novel Plasmonic Fiber Optic Biosensor Design.Plasmonics (Norwell, Mass.) · 2022Article
- Misinterpretation of solid sphere equivalent refractive index measurements and smallest detectable diameters of extracellular vesicles by flow cytometry.Scientific reports · 2021Review
- Toxicological Analysis of Hepatocytes Using FLIM Technique: In Vitro versus Ex Vivo Models.Cells · 2021Article
- Instant FLIM enables 4DOptica · 2021Article
- High-speed compressed-sensing fluorescence lifetime imaging microscopy of live cells.Proceedings of the National Academy of Sciences of the United States of America · 2021Article
- Time-Resolved Fluorescence Anisotropy and Molecular Dynamics Analysis of a Novel GFP Homo-FRET Dimer.Biophysical journal · 2021Article
- Numerical Analysis of a Highly Sensitive Surface Plasmon Resonance Sensor for SARS-CoV-2 Detection.Plasmonics (Norwell, Mass.) · 2021Article
- FCMCytometry. Part A : the journal of the International Society for Analytical Cytology · 2020Article
- Enrichment of rare events using a multi-parameter high throughput microfluidic droplet sorter.Lab on a chip · 2020Article
- Membrane Dynamics and Organization of the Phagocyte NADPH Oxidase in PLB-985 Cells.Frontiers in cell and developmental biology · 2020Article
- Fluorescence lifetime imaging reveals heterogeneous functional distribution of eGFP expressed in Xenopus oocytes.Methods and applications in fluorescence · 2019Article
- Chromatin condensation fluctuations rather than steady-state predict chromatin accessibility.Nucleic acids research · 2019Article
- Diattenuation Imaging reveals different brain tissue properties.Scientific reports · 2019Article
- Cycles of protein condensation and discharge in nuclear organelles studied by fluorescence lifetime imaging.Nature communications · 2019Article
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Authors and funding
7 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
We show that fluorescence lifetime imaging microscopy (FLIM) of green fluorescent protein (GFP) molecules in cells can be used to report on the local refractive index of intracellular GFP. We expressed GFP fusion constructs of Rac2 and gp91(phox), which are both subunits of the phagocyte NADPH oxidase enzyme, in human myeloid PLB-985 cells and showed by high-resolution confocal fluorescence microscopy that GFP-Rac2 and GFP-gp91(phox) are targeted to the cytosol and to membranes, respectively. Frequency-domain FLIM experiments on these PLB-985 cells resulted in average fluorescence lifetimes of 2.70 ns for cytosolic GFP-Rac2 and 2.31 ns for membrane-bound GFP-gp91(phox). By comparing these lifetimes with a calibration curve obtained by measuring GFP lifetimes in PBS/glycerol mixtures of known refractive index, we found that the local refractive indices of cytosolic GFP-Rac2 and membrane-targeted GFP-gp91(phox) are approximately 1.38 and approximately 1.46, respectively, which is in good correspondence with reported values for the cytosol and plasma membrane measured by other techniques. The ability to measure the local refractive index of proteins in living cells by FLIM may be important in revealing intracellular spatial heterogeneities within organelles such as the plasma and phagosomal membrane.
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