Fluorescent nanosensors for intracellular chemical analysis: decyl methacrylate liquid polymer matrix and ion-exchange-based potassium PEBBLE sensors with real-time application to viable rat C6 glioma cells
Document Type
Article
Abstract
Fluorescent spherical nanosensors, or PEBBLEs (probes encapsulated by biologically localized embedding), in the 500 nm-1 microm size range have been developed using decyl methacrylate as a matrix. A general scheme for the polymerization and introduction of sensing components creates a matrix that allows for the utilization of the highly selective ionophores used in poly(vinyl chloride) and decyl methacrylate ion-selective electrodes. We have applied these optically silent ionophores to fluorescence-based sensing by using ion-exchange and highly selective pH chromoionophores. This allows the tailoring of selective submicrometer sensors for use in intracellular measurements of important analytes for which selective enough fluorescent probes do not exist. The protocol for sensor development has been worked out for potassium sensing. It is based on the BME-44 ionophore (2-dodecyl-2-methyl-1,3-propanediylbis[N-[5'nitro(benzo-15-crown-5)-4'-yl]carbamate]). The general scheme should work for any available ionophore used in PVC or decyl methacrylate ion-selective electrodes, with minor adjustments to account for differences in ionophore charge and analyte binding constant. The reversible and highly selective sensors developed have a subsecond response time and an adjustable dynamic range. Applications to live C6 glioma cells demonstrate their utility; the intracellular potassium activity is followed in real time upon extracellular administration of kainic acid.
Medical Subject Headings
Animals; Biosensing Techniques; Calibration; Fluorescence; Glioma (chemistry, pathology); Methacrylates (chemistry); Microscopy, Confocal; Microscopy, Electron, Scanning; Polymers; Potassium (chemistry); Rats; Tumor Cells, Cultured
Publication Date
5-15-2001
Publication Title
Analytical chemistry
ISSN
0003-2700
Volume
73
Issue
10
First Page
2221
Last Page
8
PubMed ID
11393844
Digital Object Identifier (DOI)
10.1021/ac0012041
Recommended Citation
Brasuel, M; Kopelman, R; Miller, T J.; Tjalkens, R; and Philbert, M A., "Fluorescent nanosensors for intracellular chemical analysis: decyl methacrylate liquid polymer matrix and ion-exchange-based potassium PEBBLE sensors with real-time application to viable rat C6 glioma cells" (2001). Translational Neuroscience. 2523.
https://scholar.barrowneuro.org/neurobiology/2523