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Anal Bioanal Chem. 2010 Nov;398(5):1919-25. doi: 10.1007/s00216-010-4127-4. Epub 2010 Sep 2.

Application of FLIM-FIDSAM for the in vivo analysis of hormone competence of different cell types.

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  • 1Institute of Physical and Theoretical Chemistry, University of Tübingen, Auf der Morgenstelle 8, 72076 Tübingen, Germany.

Abstract

Background fluorescence derived from subcellular compartments is a major drawback in high-resolution live imaging, especially of plant cells. A novel technique for contrast enhancement of fluorescence images of living cells expressing fluorescent fusion proteins termed fluorescence intensity decay shape analysis microscopy (FIDSAM) has been recently published and is applied here to plant cells expressing wild-type levels of a low-abundant membrane protein (BRI1-EGFP), demonstrating the applicability of FIDSAM to samples exhibiting about 80% autofluorescence. Furthermore, the combination of FIDSAM and fluorescence lifetime imaging microscopy enables the simultaneous determination and quantification of different ligand-specific responses in living cells with high spatial and temporal resolution even in samples with high autofluorescence background. Correlation of different responses can be used to determine the hormone ligand competence of different cell types as demonstrated here in BRI1-EGFP-expressing root and hypocotyl cells.

PMID:
20811880
[PubMed - indexed for MEDLINE]
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