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Journal of Biomedical Optics

Detection of dual-gene expression in arteries using an optical imaging method
Author(s): Hunter H. Chen; Xiangcan Zhan; Ananda Kumar; Xiangying Du; Holly Hammond; Linzhao Cheng; Xiaoming Yang
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Paper Abstract

We evaluate the in vivo use of an optical imaging method to detect the vascular expression of green fluorescent protein (GFP) or red fluorescent protein (RFP), and to detect the simultaneous expression of GFP and RFP after transduction into arteries by a dual-promoter lentiviral vector driving their concurrent expression. This method involves using a charge-coupled device camera to detect fluorescence, a fiber optic probe to transmit light, and optical filters to distinguish each marker. In animal models, these vectors are locally delivered to target arteries, whereas the gene for a nonfluorescent cell-surface protein is transduced into contralateral arteries as the sham control. The images show distinct areas of bright fluorescence from GFP and RFP along the target arteries on excitation; no exogenous fluorescence is observed in the controls. Measured signal intensities from arteries transduced with the single- and dual-promoter vectors exceed the autofluorescence signal from the controls. Transgene expression of GFP and RFP in vivo is confirmed with confocal microscopy. We demonstrate the use of an optical imaging method to concurrently detect two distinct fluorescent proteins, potentially permitting the expression of multiple transgenes and their localization in the vasculature to be monitored.

Paper Details

Date Published: 1 November 2004
PDF: 7 pages
J. Biomed. Opt. 9(6) doi: 10.1117/1.1803842
Published in: Journal of Biomedical Optics Volume 9, Issue 6
Show Author Affiliations
Hunter H. Chen, Johns Hopkins Univ. (United States)
Xiangcan Zhan, Johns Hopkins Univ. (United States)
Ananda Kumar, Johns Hopkins Univ. (United States)
Xiangying Du, Johns Hopkins Univ. (United States)
Holly Hammond, Johns Hopkins Univ. (United States)
Linzhao Cheng, Johns Hopkins Univ. (United States)
Xiaoming Yang, Johns Hopkins Univ. (United States)

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