Authors: Nowak-Lovato, K.L.¹; Rector, K.D.²

Source: Applied Spectroscopy, Volume 63, Issue 4, Pages 96A-108A and 381-481 (April 2009) , pp. 387-395(9)

Publisher: Society for Applied Spectroscopy

Abstract:

In this paper the ability to develop a gold-core, silver-shell surface-enhanced Raman scattering (SERS) nanosensor that has both a targeting component as well as a sensing component is demonstrated. The nanosensor is capable of targeting through the FcεRI receptor-mediated endocytic pathway due to the adsorption of 2,4-ε-dinitrophenol-L-lysine (DNP) ligand on the nanoparticle surface. The nanosensor is also sensitive to pH changes in the endocytic vesicle within the pH range of 4.5-7.5 by 4-mercaptopyridine (4-MPy) adsorbed to the particle surface. The targeting and sensing moieties do not significantly interfere with each other's function. The sensing component of the nanosensor is calibrated with in vivo measurements of rat basophil leukemia (RBL-2H3) cells in standard buffer solutions using the ionophore nigericin, which serves to equilibrate the external [H⁺] concentration with that of the cell compartments. The targeting of the nanosensor is verified with a β-hexosaminidase assay. It is also demonstrated that the targeted nanosensor is capable of making accurate cellular pH measurements in RBL-2H3 cells, out to ninety minutes after targeted nanosensor addition. Whole-cell, time-lapse, hyperspectral image cubes demonstrating endocytic vesicular pH changes during FcεRI receptor mediated endocytosis are demonstrated.

Keywords: SURFACE-ENHANCED RAMAN SCATTERING; SERS; BIOSENSOR; PH SENSOR; ENDOSOMES; ENDOCYTOSIS; FCEPSIRI

Document Type: Research article

DOI: http://dx.doi.org/10.1366/000370209787944406

Affiliations: 1: Chemistry Division, Los Alamos National Laboratory, Los Alamos, New Mexico 87545; Department of Pathology, University of New Mexico, School of Medicine, Albuquerque, New Mexico 87131 2: Chemistry Division, Los Alamos National Laboratory, Los Alamos, New Mexico 87545

Publication date: 2009-04-01

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