Chapter title |
Photoactivatable Fluorescent Proteins for Super-resolution Microscopy.
|
---|---|
Chapter number | 16 |
Book title |
Photoswitching Proteins
|
Published in |
Methods in molecular biology, March 2014
|
DOI | 10.1007/978-1-4939-0470-9_16 |
Pubmed ID | |
Book ISBNs |
978-1-4939-0469-3, 978-1-4939-0470-9
|
Authors |
Yuji Ishitsuka, Karin Nienhaus, G Ulrich Nienhaus, G. Ulrich Nienhaus, Ishitsuka, Yuji, Nienhaus, Karin, Nienhaus, G. Ulrich |
Editors |
Sidney Cambridge |
Abstract |
Super-resolution fluorescence microscopy techniques such as simulated emission depletion (STED) microscopy and photoactivated localization microscopy (PALM) allow substructures, organelles or even proteins within a cell to be imaged with a resolution far below the diffraction limit of ~200 nm. The development of advanced fluorescent proteins, especially photoactivatable fluorescent proteins of the GFP family, has greatly contributed to the successful application of these techniques to live-cell imaging. Here, we will illustrate how two fluorescent proteins with different photoactivation mechanisms can be utilized in high resolution dual color PALM imaging to obtain insights into a cellular process that otherwise would not be accessible. We will explain how to set up and perform the experiment and how to use our latest software "a-livePALM" for fast and efficient data analysis. |
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