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Intracellular pH measurements using perfluorocarbon nanoemulsions
Michael J Patrick1, Jelena M Janjic, Haibing Teng
1Molecular Biosensor and Imaging Center, Carnegie Mellon University , Pittsburgh, Pennsylvania 15213, United States.
Journal of the American Chemical Society
|November 26, 2013
Summary
New perfluorocarbon (PFC) nanoemulsions with fluorescent reporters enable real-time intracellular pH measurement in living cells. This technology aids in understanding nanoemulsion fate and cell dynamics for imaging applications.
Area of Science:
- Biomedical Engineering
- Cell Biology
- Nanotechnology
Background:
- Perfluorocarbon (PFC) nanoemulsions are utilized for in vivo cell tracking via 19F MRI.
- The intracellular fate and stability of these PFC nanoemulsion probes remain poorly understood.
- Accurate intracellular pH monitoring is crucial for understanding cellular processes and probe behavior.
Purpose of the Study:
- To synthesize and characterize novel PFC nanoemulsions for intracellular pH sensing.
- To evaluate the cellular uptake, pH-sensing capabilities, and stability of these nanoemulsions.
- To explore the utility of pH-sensing PFC nanoemulsions in studying intracellular fate and cell viability.
Main Methods:
- Synthesis and formulation of PFC nanoemulsions incorporating Cy3-PFC and CypHer5-PFC fluorescent reporters.
- In vitro characterization of spectral and pH-sensing properties of the nanoemulsions.
- Longitudinal quantification of intracellular pH in rat 9L glioma cells using fluorescence microscopy and flow cytometry.
Main Results:
- Formulated PFC nanoemulsions demonstrated unaltered spectral properties of the incorporated cyanine dyes.
- Nanoemulsions were rapidly internalized by rat 9L glioma cells, localizing to acidic compartments.
- Intracellular pH was observed to decrease to approximately 5.5 within 3 hours post-incubation.
- The pH-sensing nanoemulsions enabled real-time optical detection of intracellular pH changes.
Conclusions:
- Developed PFC nanoemulsions are effective tools for real-time intracellular pH monitoring in living cells.
- These nanoemulsions facilitate the study of PFC probe intracellular fate, loading dynamics, and cell viability.
- This technology offers potential for improved understanding of cellular responses and imaging probe behavior.
