Document Detail


In vivo hyperspectral confocal fluorescence imaging to determine pigment localization and distribution in cyanobacterial cells.
MedLine Citation:
PMID:  18316743     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Hyperspectral confocal fluorescence imaging provides the opportunity to obtain individual fluorescence emission spectra in small ( approximately 0.03-microm(3)) volumes. Using multivariate curve resolution, individual fluorescence components can be resolved, and their intensities can be calculated. Here we localize, in vivo, photosynthesis-related pigments (chlorophylls, phycobilins, and carotenoids) in wild-type and mutant cells of the cyanobacterium Synechocystis sp. PCC 6803. Cells were excited at 488 nm, exciting primarily phycobilins and carotenoids. Fluorescence from phycocyanin, allophycocyanin, allophycocyanin-B/terminal emitter, and chlorophyll a was resolved. Moreover, resonance-enhanced Raman signals and very weak fluorescence from carotenoids were observed. Phycobilin emission was most intense along the periphery of the cell whereas chlorophyll fluorescence was distributed more evenly throughout the cell, suggesting that fluorescing phycobilisomes are more prevalent along the outer thylakoids. Carotenoids were prevalent in the cell wall and also were present in thylakoids. Two chlorophyll fluorescence components were resolved: the short-wavelength component originates primarily from photosystem II and is most intense near the periphery of the cell; and the long-wavelength component that is attributed to photosystem I because it disappears in mutants lacking this photosystem is of higher relative intensity toward the inner rings of the thylakoids. Together, the results suggest compositional heterogeneity between thylakoid rings, with the inner thylakoids enriched in photosystem I. In cells depleted in chlorophyll, the amount of both chlorophyll emission components was decreased, confirming the accuracy of the spectral assignments. These results show that hyperspectral fluorescence imaging can provide unique information regarding pigment organization and localization in the cell.
Authors:
Wim F J Vermaas; Jerilyn A Timlin; Howland D T Jones; Michael B Sinclair; Linda T Nieman; Sawsan W Hamad; David K Melgaard; David M Haaland
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Publication Detail:
Type:  Journal Article; Research Support, U.S. Gov't, Non-P.H.S.     Date:  2008-03-03
Journal Detail:
Title:  Proceedings of the National Academy of Sciences of the United States of America     Volume:  105     ISSN:  1091-6490     ISO Abbreviation:  Proc. Natl. Acad. Sci. U.S.A.     Publication Date:  2008 Mar 
Date Detail:
Created Date:  2008-03-12     Completed Date:  2008-04-07     Revised Date:  2010-09-22    
Medline Journal Info:
Nlm Unique ID:  7505876     Medline TA:  Proc Natl Acad Sci U S A     Country:  United States    
Other Details:
Languages:  eng     Pagination:  4050-5     Citation Subset:  IM    
Affiliation:
School of Life Sciences and Center for Bioenergy and Photosynthesis, Arizona State University, Box 874501, Tempe, AZ 85287-4501, USA.
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MeSH Terms
Descriptor/Qualifier:
Algorithms
Analysis of Variance
Biological Transport
Chlorophyll / deficiency
Microscopy, Confocal
Photosystem I Protein Complex / metabolism
Pigments, Biological / metabolism*
Spectrometry, Fluorescence
Synechocystis / cytology*,  metabolism*
Chemical
Reg. No./Substance:
0/Photosystem I Protein Complex; 0/Pigments, Biological; 1406-65-1/Chlorophyll
Comments/Corrections

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