Accuracy and Precision in Camera-Based Fluorescence Correlation Spectroscopy Measurements

• DOI: 10.1021/ac303485t
• Publication Date: 2013-03-22T20:11:41Z
• Authors: Jagadish Sankaran; Nirmalya Bag; Rachel Susan Kraut; Thorsten Wohland
• Keywords: Fluorescence Correlation Spectroscopy; Accuracy and precision; SIGNAL (programming language); Biomolecule

Imaging fluorescence correlation spectroscopy (FCS) performed using array detectors has been successfully used to quantify the number, mobility, and organization of biomolecules in cells organisms. However, there have not any systematic studies on errors these estimates that are introduced due instrumental experimental factors. State-of-the-art still restricted number frames can be recorded per unit time, sensitivity noise characteristics, total realistically recorded. These limitations place constraints time resolution, signal-to-noise ratio, measurement respectively. This work addresses problems by a combination simulations experiments lipid bilayers provide characteristic performance parameters guidelines govern accuracy precision diffusion coefficient concentration measurements camera-based FCS. We then proceed demonstrate effects capability FCS determine membrane heterogeneity via laws, showing is lower length scale limit beyond which cannot detected overcome choosing suitable parameters. On basis results, we for an efficient design extract information concentration, heterogeneity.