Simultaneous measurements of fluorescence lifetimes, anisotropy, and FRAP recovery curves
Simultaneous measurements of fluorescence lifetimes, anisotropy, and FRAP recovery curves
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同时测量荧光寿命、各向异性和 FRAP 恢复曲线
DOI:
10.1117/12.875151
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发表时间:
2011
期刊:
影响因子:
--
通讯作者:
Levitt J
中科院分区:
文献类型:
--
作者:
Levitt J
We present fluorescence lifetime imaging (FLIM) and fluorescence anisotropy imaging along with translational diffusion measurements of living cells labelled with green fluorescent protein (GFP) recorded in a single experiment. The experimental set-up allows for time and polarization-resolved fluorescence images to be measured in every frame of a fluorescence recovery after photobleaching (FRAP) series. We have validated the method using rhodamine 123 in homogeneous solution prior to measurements of living A431 cells labelled with cdc42-GFP, for which the FRAP recovery exhibits an immobile fraction and the rotational mobility of the protein is hindered while the fluorescence lifetime fairly homogeneous across the cell. By eliminating the need for sequential measurements to extract fluorescence lifetimes and molecular diffusion coefficients we remove artefacts arising from changes in sample morphology and excessive photobleaching during sequential experiments.
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DOI:
10.1021/jp066021i
发表时间:
2007
期刊:
The journal of physical chemistry. B
影响因子:
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作者:
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DOI:
--
发表时间:
2010
期刊:
影响因子:
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通讯作者:
太田信廣