Abstract
The fluorescence labeling technique is a method with a high degree of specificity and sensitivity and is often chosen as a tool in the study of protein expression and subcellular compartments (1). Recently, a large number of fluorescent dyes with distinct fluorescence excitation and emission spectra have been engineered to be used in multilabeling and co-localization experiments. Some of the most common fluorescent dyes are fluorescein isothiocyanate (FITC), Cy2, Cy3, Cy5, TRITC, and rhodamine. These dyes can be excited independently using different laser wavelengths and observed in separate fluorescent channels. The efficiency of the fluorescent probes is, however, hampered by a variable degree of spectral overlap, low quantum efficiencies and extinction coefficients, or rapid photobleaching.
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsSpringer Nature is developing a new tool to find and evaluate Protocols. Learn more
References
Di Giaimo, R., Riccio, M., Santi, S., Galeotti, C., Ambrosetti, D. C., and Melli, M. (2002) New insights into the molecular basis of progressive myoclonus epilepsy: a multiprotein complex with cystatin B. Hum. Mol. Genet. 11, 2941–2950.
Tsien, R. Y. and Waggoner, A. (1990) Fluorophores for confocal microscopy: photophysics and photochemistry, in Handbook of Biological Confocal Microscopy (Pawley, J. B., ed.), Plenum, New York, pp. 153–161.
Wells, S. and Johnson, I. (1994) Three-Dimensional Confocal Microscopy: Volume Investigation of Biological Systems (Stevens, J. K. et al., eds.), Academic Press, London U.K., pp. 101–129.
Riccio, M., Di Giaimo, R., Pianetti, S., Palmieri, P. P., Melli, M., and Santi, S. (2001). Nuclear localization of cystatin b, the cathepsin inhibitor implicated in myoclonus epilepsy (EPM1). Exp. Cell Res. 262, 84–94.
Spisni, E., Griffoni, C., Santi, S., Riccio, M., Marulli, R., Bartolini, G., et al. (2001) Colocalization prostacyclin (PGI2) synthase-caveolin-1 in endothelial cells and new roles for PGI2 in angiogenesis. Exp. Cell Res. 266, 31–43.
Manders, E. M. M., Verbeek, F. J., and Aten, J. A. (1993) Measurement of co-localization of objects in dualcolor confocal images. J. Microsc. 169, 375–382.
Tabellini, G., Bortul, R., Santi, S., Riccio, M., Baldini, G., Cappellini, A., et al. (2003) Diacylglycerol kinase-theta is localized in the speckle domains of the nucleus. Exp. Cell Res. 287, 143–154.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2004 Humana Press Inc.
About this protocol
Cite this protocol
Riccio, M., Dembic, M., Cinti, C., Santi, S. (2004). Multifluorescence Labeling and Colocalization Analyses. In: Giordano, A., Romano, G. (eds) Cell Cycle Control and Dysregulation Protocols. Methods in Molecular Biology™, vol 285. Humana Press. https://doi.org/10.1385/1-59259-822-6:171
Download citation
DOI: https://doi.org/10.1385/1-59259-822-6:171
Publisher Name: Humana Press
Print ISBN: 978-0-89603-949-0
Online ISBN: 978-1-59259-822-9
eBook Packages: Springer Protocols