Vital fluorescent labeling for confocal scanning microscopic study of living cell invasion

Allan Z. Wang; Jian M. Chen; Gregory W. Fisher; Jane C. Wang

doi:10.1117/12.277755

7 July 1997 Vital fluorescent labeling for confocal scanning microscopic study of living cell invasion

Allan Z. Wang, Jian M. Chen, Gregory W. Fisher, Jane C. Wang

Proceedings Volume 3131, Optical Scanning Systems: Design and Applications; (1997) https://doi.org/10.1117/12.277755
Event: Optical Science, Engineering and Instrumentation '97, 1997, San Diego, CA, United States

Abstract

Invasion by cells with malignant or transformed phenotypes precedes destruction of adjacent tissue and fatal cell metastasis. State-of-the-art confocal laser scanning technology facilitates both in vitro and in vivo research into cell invasion and metastasis. In particular, studies performed with living cells yield more precise information than those with fixed cells, giving new insight into cell invasion and metastasis. We have tested a variety of vital florescent dyes and fluorogenic protease substrates in our studies of invasion of cartilage by transformed synoviocytes or osteosarcoma cells. The fluorescent dyes tested include Calcein acetoxy methyl-FITC (Calcein), Hoechst 33342 (Hoechst), CellTracker, DiI, DiO, DiD, and ethidium bromide (EB). The fluorogenic protease substrate used Meoxysuccinyl-Gly-Pro-Leu-Gly-Pro-AFC (MOS-GPLGP-AFC) for detection of collagenase activity. We found that Calcein-FITC labeling permitted the clearest direct observation of the penetration of transformed synoviocytes and osteosarcoma cells into cartilage. Even better results were obtained when chondrocyte nuclei were counter-stained with Hoechst 33342. During the invasion process, collagenase activity was observed around the synoviocyte in the cartilage matrix labeled with the fluorogenic collagenase substrate. We concluded that of the vital fluorescent dyes tested, a combined application of Calcein-FITC, Hoechst 23223, and MOS- GPLGP-AFC is most appropriate for the study of the cell invasion process.

Citation Download Citation

Allan Z. Wang, Jian M. Chen, Gregory W. Fisher, and Jane C. Wang "Vital fluorescent labeling for confocal scanning microscopic study of living cell invasion", Proc. SPIE 3131, Optical Scanning Systems: Design and Applications, (7 July 1997); https://doi.org/10.1117/12.277755

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KEYWORDS

Cartilage

Confocal microscopy

Luminescence

In vitro testing

Tissues

Tumors

In vivo imaging

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