A Simple Method for GFP- and RFP-based Dual Color Single-Molecule Localization Microscopy
| Autor: | Evgenia Platonova, Helge Ewers, Christian M. Winterflood |
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| Rok vydání: | 2015 |
| Předmět: |
Single molecule localization
Recombinant Fusion Proteins Caveolin 1 Green Fluorescent Proteins Color Nanotechnology Biochemistry Green fluorescent protein Cell Line Tumor Chromatic aberration Microscopy Humans Nanoscopic scale Membrane Glycoproteins Osteoblasts Chemistry Resolution (electron density) General Medicine Single-Domain Antibodies Nuclear Pore Complex Proteins Luminescent Proteins Gene Expression Regulation Microscopy Fluorescence Molecular Medicine Dual color Signal Transduction |
| Zdroj: | ACS Chemical Biology. 10:1411-1416 |
| ISSN: | 1554-8937 1554-8929 |
| DOI: | 10.1021/acschembio.5b00046 |
| Popis: | The recent development of single-molecule localization-based super-resolution techniques has afforded a resolution in the nanometer range in light microscopy. The ability to resolve biological structures on this scale by multicolor techniques faces significant challenges which have prevented their widespread use. Here, we provide a generic approach for high-quality simultaneous two-color single-molecule localization microscopy imaging of any combination of GFP- and RFP-tagged proteins with the use of nanobodies. Our method addresses a number of common issues related to two-color experiments, including accuracy and density of labeling as well as chromatic aberration and color-crosstalk with only minimal technical requirements. We demonstrate two-color imaging of various nanoscopic structures and show a compound resolution down to the limit routinely achieved only in a single color. |
| Databáze: | OpenAIRE |
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