Share Email Print

Proceedings Paper

Multipixel spectral imaging of green fluorescent protein (GFP) in COS-7 cells: folding kinetics and chromophore formation
Author(s): Lior Greenbaum; Chana Rothmann; Judith Hanania; Zvi Malik
Format Member Price Non-Member Price
PDF $17.00 $21.00

Paper Abstract

Spectrally resolved imaging of Green fluorescent protein (GFP) expressed in living COS-7 kidney cells distinguished the subcellular localization and demarcated the processes of protein folding and chromophore formation. COS-7 kidney cells were transfected by a plasmid pEGFP-N1 plasmid followed by incubation for 15 hours for gen expression. At different intervals the cells were examined by fluorescence microscopy and analyzed by a spectral imaging system. After 7 hours, GFP was detected in the cytoplasm, concentrated in a localized form. Spectra of the initial GFP evinced several components that belong both tot he typical fluorescent signal as well as to unspecific fingerprints. At 10 and 15 hours, GFP was seen spread in the cytoplasm as well as in the nucleus, and the specific spectra of the GFP were dominant at the later time. The typical GFP spectral fingerprint is the result of protein folding and chromophore formation following internal oxidation reactions. This folding and chromophore formation process, up to final conformation, was detected by spectral imaging as localized in the nucleus rather than in the cytosol. Thus, the method of fluorescence microscopy combined with multiplex spectral imaging demonstrates distinct biochemical pathways leading to GFP conformation.

Paper Details

Date Published: 18 December 2000
PDF: 5 pages
Proc. SPIE 4164, Laser Microscopy, (18 December 2000); doi: 10.1117/12.410631
Show Author Affiliations
Lior Greenbaum, Bar-Ilan Univ. (Israel)
Chana Rothmann, Bar-Ilan Univ. (Israel)
Judith Hanania, Bar-Ilan Univ. (Israel)
Zvi Malik, Bar-Ilan Univ. (Israel)

Published in SPIE Proceedings Vol. 4164:
Laser Microscopy
Karsten Koenig; Hans J. Tanke; Herbert Schneckenburger, Editor(s)

© SPIE. Terms of Use
Back to Top
Sign in to read the full article
Create a free SPIE account to get access to
premium articles and original research
Forgot your username?