Expression of GFP in tumor cells and fluorescent examination by confocal microscope

Ying Jin; Da Xing; Chaoyang Xu

doi:10.1117/12.462552

12 April 2002 Expression of GFP in tumor cells and fluorescent examination by confocal microscope

Ying Jin, Da Xing, Chaoyang Xu

Proceedings Volume 4536, International Workshop on Photonics and Imaging in Biology and Medicine; (2002) https://doi.org/10.1117/12.462552
Event: International Workshop on Photonics and Imaging in Biology and Medicine, 2001, Wuhan, China

Abstract

The green fluorescent protein (GFP), from the bioluminescent jellyfish Aequorea victoria, yields a bright green fluorescence when expressed in either eukaryotic or prokaryotic cells and illuminated by blue or UV light. The characteristic properties of GFP make this protein a good candidate for use as a molecular reporter to monitor patterns of protein localization, gene expression, and intracellular protein trafficking in living cells. In this study, the plasmid EGFP encoding GFP was used to transfect SWO cells (a cancer cell line of nerve gelatinous tissue) mediated by liposome: (1) The plasmid EGFP-C1, purchased from Clontech Co., propagated in suitable E. coli strain (JM 109), was extracted by Concert High Purity Plasmid Miniprep (Gibco). (2) SWO was cultured in RPMI 1640 (10% FCS and 25 mM HEPES), 37 degree(s)C, 5% CO₂. Cancer cells were transfected in 6-cm tissue culture dishes by Lipofectin Reagent (Gibco) for 6-12 hr using 2 ug DNA. (3) Then, infected cells were collected in medium containing 800 ug/ml G418, and the resistant clones were harvested and subcloned in fresh culture medium maintaining 800 ug/ml G418. (4) The cells were examined by using Nikon fluorescent microscope (E600) and Bio-Rad confocal microscope (MRC 600). (5) Next step, the cancer cells, stably expressing GFP after in vivo transduction, were implanted by surgical orthotopic implantation (SOI) in nude mice. Tracking of these cancer cells will become more sensitive and rapid than the traditional procedure of histopathological examination or immunohistochemistry. This method demonstrates external, noninvasive, whole-body, real-time fluorescence optical imaging of internally growing tumors and metastases in transplanted animals.

Citation Download Citation

Ying Jin, Da Xing, and Chaoyang Xu "Expression of GFP in tumor cells and fluorescent examination by confocal microscope", Proc. SPIE 4536, International Workshop on Photonics and Imaging in Biology and Medicine, (12 April 2002); https://doi.org/10.1117/12.462552

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