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Journal of Biomedical Optics

Quantification of endocytosis using a folate functionalized silica hollow nanoshell platform
Author(s): Sergio Sandoval; Natalie Mendez; Jesus G. Alfaro; Jian Yang; Sharraya Aschemeyer; Alexander Liberman; William C. Trogler; Andrew C. Kummel
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Paper Abstract

A quantification method to measure endocytosis was designed to assess cellular uptake and specificity of a targeting nanoparticle platform. A simple N-hydroxysuccinimide ester conjugation technique to functionalize 100-nm hollow silica nanoshell particles with fluorescent reporter fluorescein isothiocyanate and folate or polyethylene glycol (PEG) was developed. Functionalized nanoshells were characterized using scanning electron microscopy and transmission electron microscopy and the maximum amount of folate functionalized on nanoshell surfaces was quantified with UV-Vis spectroscopy. The extent of endocytosis by HeLa cervical cancer cells and human foreskin fibroblast (HFF-1) cells was investigated in vitro using fluorescence and confocal microscopy. A simple fluorescence ratio analysis was developed to quantify endocytosis versus surface adhesion. Nanoshells functionalized with folate showed enhanced endocytosis by cancer cells when compared to PEG functionalized nanoshells. Fluorescence ratio analyses showed that 95% of folate functionalized silica nanoshells which adhered to cancer cells were endocytosed, while only 27% of PEG functionalized nanoshells adhered to the cell surface and underwent endocytosis when functionalized with 200 and 900  μg, respectively. Additionally, the endocytosis of folate functionalized nanoshells proved to be cancer cell selective while sparing normal cells. The developed fluorescence ratio analysis is a simple and rapid verification/validation method to quantify cellular uptake between datasets by using an internal control for normalization.

Paper Details

Date Published: 28 August 2015
PDF: 15 pages
J. Biomed. Opt. 20(8) 088003 doi: 10.1117/1.JBO.20.8.088003
Published in: Journal of Biomedical Optics Volume 20, Issue 8
Show Author Affiliations
Sergio Sandoval, Univ. of California, San Diego (United States)
Natalie Mendez, Univ. of California, San Diego (United States)
Jesus G. Alfaro, Univ. of California, San Diego (United States)
Jian Yang, Univ. of California, San Diego (United States)
Sharraya Aschemeyer, Univ. of California, San Diego (United States)
Alexander Liberman, Univ. of California, San Diego (United States)
William C. Trogler, Univ. of California, San Diego (United States)
Andrew C. Kummel, Univ. of California, San Diego (United States)


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