Special Section on Nanophotonics for Diagnostics, Protection, and Treatment of Cancer and Inflammatory Diseases

Delivery of optical contrast agents using Triton-X100, part 1: reversible permeabilization of live cells for intracellular labeling

[+] Author Affiliations
Anne L. van de Ven

Rice University, Department of Bioengineering, MS 142, Keck Hall, Suite 116, 6100 Main Street, Houston, Texas 77005

Karen Adler-Storthz

University of Texas, Dental Branch, Department of Diagnostic Sciences, 6516 M.D. Anderson Blvd., DBB, 4.133, Houston, Texas 77030

Rebecca Richards-Kortum

Rice University, Department of Bioengineering, MS 142, Keck Hall, Suite 116, 6100 Main Street, Houston, Texas 77005

J. Biomed. Opt. 14(2), 021012 (March 10, 2009). doi:10.1117/1.3090448
History: Received August 04, 2008; Revised November 11, 2008; Accepted December 09, 2008; Published March 10, 2009
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Effective delivery of optical contrast agents into live cells remains a significant challenge. We sought to determine whether Triton-X100, a detergent commonly used for membrane isolation and protein purification, could be used to effectively and reversibly permeabilize live cells for delivery of targeted optical contrast agents. Although Triton-X100 is widely recognized as a good cell permeabilization agent, no systematic study has evaluated the efficiency, reproducibility, and reversibility of Triton-X100–mediated permeabilization in live mammalian cells. We report a series of studies to characterize macromolecule delivery in cells following Triton-X100 treatment. Using this approach, we demonstrate that molecules ranging from 1to150kDa in molecular weight can be reproducibly delivered into live cells by controlling the moles of Triton-X100 relative to the number of cells to be treated. When Triton-X100 is administered at or near the minimum effective concentration, cell permeabilization is generally reversed within 24h, and treated cells continue to proliferate and show metabolic activity during the restoration of membrane integrity. We conclude that Triton-X100 is a promising permeabilization agent for efficient and reproducible delivery of optical contrast agents into live mammalian cells.

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© 2009 Society of Photo-Optical Instrumentation Engineers

Citation

Anne L. van de Ven ; Karen Adler-Storthz and Rebecca Richards-Kortum
"Delivery of optical contrast agents using Triton-X100, part 1: reversible permeabilization of live cells for intracellular labeling", J. Biomed. Opt. 14(2), 021012 (March 10, 2009). ; http://dx.doi.org/10.1117/1.3090448


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