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Liquid polymer nano-PEBBLEs for Cl- analysis and biological applications
Murphy G Brasuel1, Terry J Miller, Raoul Kopelman
1Department of Chemistry, University of Michigan, Ann Arbor, MI 48109-1055, USA.
The Analyst
|December 12, 2003
Summary
Researchers developed novel nanometer-scale fluorescent nanosensors, called PEBBLEs (probes encapsulated by biologically localized embedding), for intracellular chloride monitoring. Method 3 demonstrated a low detection limit and linear dynamic range, enabling real-time cellular measurements.
Area of Science:
- Nanotechnology
- Biomedical Engineering
- Analytical Chemistry
Background:
- Intracellular anion monitoring is crucial for understanding cellular processes.
- Existing fluorescent probes lack selectivity for biologically relevant anions.
- Submicron sensors are needed for precise intracellular measurements.
Purpose of the Study:
- To develop the first nanometer-scale anion-sensing fluorescent spherical nanosensors (PEBBLEs) for intracellular chloride monitoring.
- To adapt existing sensor schemes for selective anion detection.
- To enable real-time monitoring of intracellular chloride levels in cells.
Main Methods:
- Developed PEBBLEs using a polymerization scheme incorporating selective ionophores and chromoionophores.
- Investigated three methods for chloride-sensitive PEBBLEs, focusing on ionophore-chromoionophore combinations.
- Utilized Method 3 (Chloride ionophore III and chromoionophore III) for indirect Cl- activity monitoring via H+ coextraction.
- Delivered PEBBLEs into C6 glioma cells using a gene gun for in vivo measurements.
Main Results:
- Method 3 yielded the most promising results for chloride-sensitive PEBBLEs.
- Achieved a limit of detection of 0.2 mM Cl- at pH 7.2.
- Established a linear dynamic range of 0.4 mM-190 mM Cl-.
- Successfully monitored intracellular chloride levels in C6 glioma cells during kainic acid stimulation.
Conclusions:
- The developed PEBBLEs represent a significant advancement in intracellular anion sensing.
- Method 3 offers a viable approach for selective and sensitive intracellular chloride monitoring.
- These nanosensors facilitate real-time studies of cellular ion channel activity and related physiological processes.