13 February 2009 Adding new dimensions to fluorescence microscopy
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Global analysis algorithms perform better than unconstrained data fitting and can improve the accuracy and precision of the experimental data. Here, we report on different strategies that can be pursued to improve the results derived from fluorescence measurements. We point out the benefits of acquiring fluorescence data in a temporally and spectrally resolved manner and show how these data sets can be used to evaluate FRET measurements. Fluorescence measurements can be also combined with other methods such as the patch-clamp technique. This combination allows to record simultaneously fluorescence signals and electrical currents of ion channels in membrane patches. Global analysis of the data can yield valuable information about the processes underlying channel activation. We used this approach to study the activation of homotetrameric CNGA2 channels in inside-out membrane patches. By using fluorescent analogues of cyclic nucleotides as ligands we were able to simultaneously determine ligand binding and channel activation.
© (2009) COPYRIGHT Society of Photo-Optical Instrumentation Engineers (SPIE). Downloading of the abstract is permitted for personal use only.
Christoph Biskup, Christoph Biskup, Jana Kusch, Jana Kusch, Eckhard Schulz, Eckhard Schulz, Birgit Hoffmann, Birgit Hoffmann, Vasilica Nache, Vasilica Nache, Frank Schwede, Frank Schwede, Frank Lehmann, Frank Lehmann, Klaus Benndorf, Klaus Benndorf, } "Adding new dimensions to fluorescence microscopy", Proc. SPIE 7183, Multiphoton Microscopy in the Biomedical Sciences IX, 718308 (13 February 2009); doi: 10.1117/12.809706; https://doi.org/10.1117/12.809706


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