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Journal of Sensors
Volume 2009, Article ID 235158, 4 pages
http://dx.doi.org/10.1155/2009/235158
Research Article

Characterization of PEBBLEs as a Tool for Real-Time Measurement of Dictyostelium discoideum Endosomal pH

1Department of Chemistry/Biochemistry, Colorado College, 14 East Cache La Poudre Street, Colorado Springs, CO 80903, USA
2Department of Biology, Colorado College, 14 East Cache La Poudre Street, Colorado Springs, CO 80903, USA

Received 26 May 2009; Revised 21 July 2009; Accepted 4 August 2009

Academic Editor: Mike McShane

Copyright © 2009 Everett Moding et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

The measurement of intracellular ion concentration change is important for understanding the cellular mechanisms for communication. Recently developed nanosensors, (Photonic Explorers for Biomedical use with Biologically Localized Embedding) PEBBLEs, have a number of advantages for measuring ions in cells over established methods using microelectrodes, unbound fluorescent dyes, or NMR. PEBBLE sensors have been shown to work in principle for measuring dynamic ion changes, but few in vivo applications have been demonstrated. We modified the protocol for the fabrication of pH sensing PEBBLEs and developed a protocol for the utilization of these sensors for the monitoring of dynamic pH changes in the endosomes of slime mold Dictyostelium discoideum (D. discoideum). Oregon Green 514-CdSe Quantum Dot PEBBLEs were used to measure real-time pH inside D. discoideum endosomes during cAMP stimulation. Endosomal pH was shown to decrease during cAMP signaling, demonstrating a movement of protons into the endosomes of D. discoideum amoebae.