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Single-Molecule Tracking Microscopy - A Tool for Determining the Diffusive States of Cytosolic Molecules
Published on: September 5, 2019
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Accounting for limited detection efficiency and localization precision in cluster analysis in single molecule
Arun Shivanandan1, Jayakrishnan Unnikrishnan2, Aleksandra Radenovic1
1Laboratory of Nanoscale Biology, Institute of Bioengineering, Ecole Polytechnique Federale de Lausanne (EPFL), Lausanne, Switzerland.
Plos One
|March 21, 2015
Summary
Single Molecule Localization Microscopy (SMLM) methods can now accurately quantify membrane protein organization. New analytical techniques correct for detection and localization errors, improving cluster analysis in quantitative biology.
Area of Science:
- Biophysics
- Cell Biology
- Microscopy
Background:
- Single Molecule Localization Microscopy (SMLM) techniques, such as PhotoActivated Localization Microscopy, offer sub-diffraction limit resolution for studying membrane protein spatial organization.
- Quantitative cluster analysis using SMLM is popular but limited by inherent errors.
Purpose of the Study:
- To develop analytical methods for estimating and correcting errors in SMLM-based quantitative cluster analysis.
- To improve the accuracy and precision of membrane protein spatial organization quantification.
Main Methods:
- Utilized Ripley's L(r) - r and Pair Correlation Function for cluster analysis.
- Developed analytical methods to estimate and correct for SMLM errors, including detection efficiency and localization precision.
Main Results:
- Provided analytical methods to address SMLM's limitations in detection efficiency (<60%) and localization precision (10-30 nm).
- Demonstrated a way to correct for errors affecting quantitative cluster analysis.
Conclusions:
- The developed methods offer a more accurate and precise quantification of protein spatial organization.
- These advancements can lead to breakthrough results in quantitative biology by refining SMLM data analysis.
