Needham, SR and Hirsch, M and Rolfe, DJ and Clarke, DT and Zanetti-Domingues, LC and Wareham, R and Martin-Fernandez, ML (2013) Measuring EGFR Separations on Cells with ∼10 nm Resolution via Fluorophore Localization Imaging with Photobleaching. PLoS ONE, 8.Full text not available from this repository.
Detecting receptor dimerisation and other forms of clustering on the cell surface depends on methods capable of determining protein-protein separations with high resolution in the ∼10-50 nm range. However, this distance range poses a significant challenge because it is too large for fluorescence resonance energy transfer and contains distances too small for all other techniques capable of high-resolution in cells. Here we have adapted the technique of fluorophore localisation imaging with photobleaching to measure inter-receptor separations in the cellular environment. Using the epidermal growth factor receptor, a key cancer target molecule, we demonstrate ∼10 nm resolution while continuously covering the range of ∼10-80 nm. By labelling the receptor on cells expressing low receptor numbers with a fluorescent antagonist we have found inter-receptor separations all the way up from 8 nm to 59 nm. Our data are consistent with epidermal growth factor receptors being able to form homo-polymers of at least 10 receptors in the absence of activating ligands. © 2013 Needham et al.
|Divisions:||Div F > Signal Processing and Communications|
|Depositing User:||Cron Job|
|Date Deposited:||15 Dec 2015 12:55|
|Last Modified:||06 May 2016 06:44|