Normalized Polarization Ratios for the Analysis of Cell Polarity
Raz Shimoni, Kim Pham, Mohammed Yassin, Mandy J Ludford-Menting, Min Gu, Sarah M Russell
PLoS One | PUBLIC LIBRARY SCIENCE | Published : 2014
The quantification and analysis of molecular localization in living cells is increasingly important for elucidating biological pathways, and new methods are rapidly emerging. The quantification of cell polarity has generated much interest recently, and ratiometric analysis of fluorescence microscopy images provides one means to quantify cell polarity. However, detection of fluorescence, and the ratiometric measurement, is likely to be sensitive to acquisition settings and image processing parameters. Using imaging of EGFP-expressing cells and computer simulations of variations in fluorescence ratios, we characterized the dependence of ratiometric measurements on processing parameters. This a..View full abstract
Awarded by National Health and Medical Research Foundation
Awarded by Australian Research Council
Awarded by Human Frontiers Science Program
This work was supported by the National Health and Medical Research Foundation, https://www.nhmrc.gov.au/, 1020234, 620500; the Australian Research Council, http://www.arc.gov.au/, FT0990405; and the Human Frontiers Science Program, http://www.hfsp.org/, RGP0049/2007. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.