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Optical Nanosensors for Chemical Analysis inside Single Living Cells. 2. Sensors for pH and Calcium and the Intracellular Application of PEBBLE Sensors

Optical nanosensors, or PEBBLEs (probes encapsulated by biologically localized embedding), have been produced for intracellular measurements of pH and calcium. Five varieties of pH-sensitive sensors and three different calcium-selective sensors are presented and discussed. Each sensor combines an io...

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Published in:Analytical chemistry (Washington) 1999-11, Vol.71 (21), p.4837-4843
Main Authors: Clark, Heather A, Kopelman, Raoul, Tjalkens, Ron, Philbert, Martin A
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creator Clark, Heather A
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Tjalkens, Ron
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description Optical nanosensors, or PEBBLEs (probes encapsulated by biologically localized embedding), have been produced for intracellular measurements of pH and calcium. Five varieties of pH-sensitive sensors and three different calcium-selective sensors are presented and discussed. Each sensor combines an ion-selective fluorescent indicator and an ion-insensitive internal standard entrapped within an acrylamide polymeric matrix. Calibrations and linear ranges are presented for each sensor. The photobleaching of dyes incorporated into PEBBLEs is comparable to that of the respective free dye that is incorporated within the matrix. These PEBBLE sensors are fully reversible over many measurements. The leaching of fluorescent indicator from the polymer is less than 50% over a 48-h period (note that a typical application time is only a few hours). The PEBBLE sensors have also been applied to intracellular analysis of the calcium flux in the cytoplasm of neural cells during the mitochondrial permeability transition. Specifically, a distinct difference is noted between cells of different types (astrocyte vs neuron-derived cells) with respect to their response to the toxicant m-dinitrobenzene (DNB). Use of PEBBLE sensors permits the quantitative discrimination of subtle differences between the ability of human SY5Y neuroblastoma and C6 glioma to respond to challenge with DNB. Specifically, measurement of intracellular calcium, the precursor to cell death, has been achieved.
doi_str_mv 10.1021/ac990630n
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Sensors for pH and Calcium and the Intracellular Application of PEBBLE Sensors</atitle><jtitle>Analytical chemistry (Washington)</jtitle><addtitle>Anal. Chem</addtitle><date>1999-11-01</date><risdate>1999</risdate><volume>71</volume><issue>21</issue><spage>4837</spage><epage>4843</epage><pages>4837-4843</pages><issn>0003-2700</issn><eissn>1520-6882</eissn><coden>ANCHAM</coden><abstract>Optical nanosensors, or PEBBLEs (probes encapsulated by biologically localized embedding), have been produced for intracellular measurements of pH and calcium. Five varieties of pH-sensitive sensors and three different calcium-selective sensors are presented and discussed. Each sensor combines an ion-selective fluorescent indicator and an ion-insensitive internal standard entrapped within an acrylamide polymeric matrix. Calibrations and linear ranges are presented for each sensor. The photobleaching of dyes incorporated into PEBBLEs is comparable to that of the respective free dye that is incorporated within the matrix. These PEBBLE sensors are fully reversible over many measurements. The leaching of fluorescent indicator from the polymer is less than 50% over a 48-h period (note that a typical application time is only a few hours). The PEBBLE sensors have also been applied to intracellular analysis of the calcium flux in the cytoplasm of neural cells during the mitochondrial permeability transition. Specifically, a distinct difference is noted between cells of different types (astrocyte vs neuron-derived cells) with respect to their response to the toxicant m-dinitrobenzene (DNB). Use of PEBBLE sensors permits the quantitative discrimination of subtle differences between the ability of human SY5Y neuroblastoma and C6 glioma to respond to challenge with DNB. Specifically, measurement of intracellular calcium, the precursor to cell death, has been achieved.</abstract><cop>Washington, DC</cop><pub>American Chemical Society</pub><pmid>10565275</pmid><doi>10.1021/ac990630n</doi><tpages>7</tpages></addata></record>
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subjects Biological and medical sciences
Calcium
Calcium - analysis
Calibration
Cancer
Cell Membrane Permeability - drug effects
Cells
Chemistry
Coloring Agents - chemistry
Dinitrobenzenes - pharmacology
Diverse techniques
Fluoresceins - chemistry
Fluorescent Dyes - chemistry
Fundamental and applied biological sciences. Psychology
Glioma - drug therapy
Glioma - metabolism
Humans
Hydrogen-Ion Concentration
Intracellular Membranes - drug effects
Microscopy, Confocal - methods
Mitochondria - drug effects
Mitochondria - metabolism
Molecular and cellular biology
Neuroblastoma - drug therapy
Neuroblastoma - metabolism
Optics
Optics and Photonics - instrumentation
Organic Chemicals
Photochemistry - methods
Prostheses and Implants
Sensors
Tumor Cells, Cultured
title Optical Nanosensors for Chemical Analysis inside Single Living Cells. 2. Sensors for pH and Calcium and the Intracellular Application of PEBBLE Sensors
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