Adding new dimensions to fluorescence microscopy

Christoph Biskup; Jana Kusch; Eckhard Schulz; Birgit Hoffmann; Vasilica Nache; Frank Schwede; Frank Lehmann; Klaus Benndorf

doi:10.1117/12.809706

13 February 2009 Adding new dimensions to fluorescence microscopy

Christoph Biskup, Jana Kusch, Eckhard Schulz, Birgit Hoffmann, Vasilica Nache, Frank Schwede, Frank Lehmann, Klaus Benndorf

Author Affiliations +

Proceedings Volume 7183, Multiphoton Microscopy in the Biomedical Sciences IX; 718308 (2009) https://doi.org/10.1117/12.809706
Event: SPIE BiOS, 2009, San Jose, California, United States

Abstract

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.

Citation Download Citation

Christoph Biskup, Jana Kusch, Eckhard Schulz, Birgit Hoffmann, Vasilica Nache, Frank Schwede, Frank Lehmann, and Klaus Benndorf "Adding new dimensions to fluorescence microscopy", Proc. SPIE 7183, Multiphoton Microscopy in the Biomedical Sciences IX, 718308 (13 February 2009); https://doi.org/10.1117/12.809706

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KEYWORDS

Luminescence

Data modeling

Confocal microscopy

Microscopes

Fluorescence resonance energy transfer

Microscopy

Photons

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