ArticleScientific reports2021
Label-free characterization of single extracellular vesicles using two-photon fluorescence lifetime imaging microscopy of NAD(P)H.
Article in Scientific reports, 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 19 papers.
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Who cites it
19 citing papers in PubMed.
- Emerging trends of fluorescence lifetime imaging microscopy (FLIM): advances, challenges, and prospects.Biophysics reports · 2026Article
- On the importance of simultaneous label-free multimodal nonlinear optical imaging for biomedical applications.APL photonics · 2025Article
- Spatial Distribution of Extracellular Vesicles, Autofluorescence and CD9 Positivity Around Chondrocytes in the Superficial Layer of Articular Cartilage.Journal of extracellular vesicles · 2025Article
- Adipose Tissue-Derived Extracellular Vesicle MicroRNAs: Diagnostic Biomarkers for the Pathophysiology Associated with Obesity.Precision chemistry · 2025Review
- Extracellular Vesicles for Clinical Diagnostics: From Bulk Measurements to Single-Vesicle Analysis.ACS nano · 2025Review
- Label-free method to monitor metabolism during long-term culture of human pluripotent stem cell derived cardiomyocytes.Biophotonics discovery · 2025Article
- Modeling the pre-metastatic niche of gastric cancer peritoneal metastasis under spatiotemporal resolution and investigating EVs-mediated immune suppression.Frontiers in immunology · 2025Review
- Label-free nonlinear optical signatures of extracellular vesicles in liquid and tissue biopsies of human breast cancer.Scientific reports · 2024Article
- Label-free approaches for extracellular vesicle detection.iScience · 2023Review
- Formalin fixation and paraffin embedding interfere with the preservation of optical metabolic assessments based on endogenous NAD(P)H and FAD two-photon excited fluorescence.Biomedical optics express · 2023Article
- In vivo label-free optical signatures of chemotherapy response in human pancreatic ductal adenocarcinoma patient-derived xenografts.Communications biology · 2023Article
- A Review of Labeling Approaches Used in Small Extracellular Vesicles Tracing and Imaging.International journal of nanomedicine · 2023Review
- Isolation and characterization of extracellular vesicles and future directions in diagnosis and therapy.Wiley interdisciplinary reviews. Nanomedicine and nanobiotechnology · 2023Review
- A Criterion of Colorectal Cancer Diagnosis Using Exosome Fluorescence-Lifetime Imaging.Diagnostics (Basel, Switzerland) · 2022Article
- Changing Perspectives from Oxidative Stress to Redox Signaling-Extracellular Redox Control in Translational Medicine.Antioxidants (Basel, Switzerland) · 2022Review
- Label-free optical redox ratio from urinary extracellular vesicles as a screening biomarker for bladder cancer.American journal of cancer research · 2022Article
- Megahertz scan rates enabled by optical sampling by repetition-rate tuning.Scientific reports · 2021Article
- Real-time pixelwise phasor analysis for video-rate two-photon fluorescence lifetime imaging microscopy.Biomedical optics express · 2021Article
- Label-Free Multiphoton Microscopy for the Detection and Monitoring of Calcific Aortic Valve Disease.Frontiers in cardiovascular medicine · 2021Review
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Authors and funding
8 authors.
Funding
Abstract
The heterogeneous nature of extracellular vesicles (EVs) creates the need for single EV characterization techniques. However, many common biochemical and functional EV analysis techniques lack single EV resolution. Two-photon fluorescence lifetime imaging microscopy (FLIM) is widely used to functionally characterize the reduced form of nicotinamide adenine dinucleotide and nicotinamide adenine dinucleotide phosphate (NAD(P)H) in cells and tissues. Here, we demonstrate that FLIM can also be used to image and characterize NAD(P)H in single isolated EVs. EVs were isolated using standard differential ultracentrifugation techniques from multiple cell lines and imaged using a custom two-photon FLIM system. The presented data show that the NAD(P)H fluorescence lifetimes in isolated cell-derived EVs follow a wide Gaussian distribution, indicating the presence of a range of different protein-bound and free NAD(P)H species. EV NAD(P)H fluorescence lifetime distribution has a larger standard deviation than that of cells and a significantly different fluorescence lifetime distribution than the nuclei, mitochondria, and cytosol of cells. Additionally, changes in the metabolic conditions of cells were reflected in changes in the mean fluorescence lifetime of NAD(P)H in the produced EVs. These data suggest that FLIM of NAD(P)H could be a valuable tool for EV research.
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