Using Photo Activated Localization Microscopy to Measure Protein Concentrations
Author Information
Author(s): Annibale Paolo, Vanni Stefano, Scarselli Marco, Rothlisberger Ursula, Radenovic Aleksandra
Primary Institution: Ecole Polytechnique Fédérale de Lausanne, Lausanne, Switzerland
Hypothesis
How can photophysical information improve quantitative measurements in PALM imaging?
Conclusion
The study presents a method to accurately estimate the number of photoblinking molecules in a sample using PALM imaging.
Supporting Evidence
- The method allows simultaneous imaging and concentration estimation over a wide expression range.
- The study validates the method by applying it to measure β2 adrenergic receptors in HeLa cells.
- The results show that the counts vs. dark time curve retains an exponential decay, confirming the distribution of off-times.
Takeaway
This study shows how scientists can count tiny proteins in cells by using special light techniques that track how these proteins blink.
Methodology
The study used PALM imaging to analyze the effects of fluorophore photoblinking on protein counting.
Limitations
The method may not account for extreme photophysical behaviors of a small fraction of molecules.
Digital Object Identifier (DOI)
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