Nanometric molecular separation measurements by single molecule photobleaching

doi:10.1016/j.ymeth.2015.05.009

. 2015 Oct 15:88:76-80.

doi: 10.1016/j.ymeth.2015.05.009. Epub 2015 May 14.

Nanometric molecular separation measurements by single molecule photobleaching

Affiliations

¹ Central Laser Facility, Research Complex at Harwell, STFC Rutherford Appleton Laboratory, Harwell Campus, Didcot OX11 0FA, UK.
² Central Laser Facility, Research Complex at Harwell, STFC Rutherford Appleton Laboratory, Harwell Campus, Didcot OX11 0FA, UK. Electronic address: daniel.rolfe@stfc.ac.uk.

PMID: 25980369
DOI: 10.1016/j.ymeth.2015.05.009

Nanometric molecular separation measurements by single molecule photobleaching

Stephen E D Webb et al. Methods. 2015.

. 2015 Oct 15:88:76-80.

doi: 10.1016/j.ymeth.2015.05.009. Epub 2015 May 14.

Affiliations

¹ Central Laser Facility, Research Complex at Harwell, STFC Rutherford Appleton Laboratory, Harwell Campus, Didcot OX11 0FA, UK.
² Central Laser Facility, Research Complex at Harwell, STFC Rutherford Appleton Laboratory, Harwell Campus, Didcot OX11 0FA, UK. Electronic address: daniel.rolfe@stfc.ac.uk.

PMID: 25980369
DOI: 10.1016/j.ymeth.2015.05.009

Abstract

Although considerable progress has been made in imaging distances in cells below the diffraction limit using FRET and super-resolution microscopy, methods for determining the separation of macromolecules in the 10-50 nm range have been elusive. We have developed fluorophore localisation imaging with photobleaching (FLImP), based on the quantised bleaching of individual protein-bound dye molecules, to quantitate the molecular separations in oligomers and nanoscale clusters. We demonstrate the benefits of using our method in studying the nanometric organisation of the epidermal growth factor receptor in cells.

Keywords: Dimerisation; Epidermal growth factor receptor; Oligomerisation; Single-molecule localisation.

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Nanometric molecular separation measurements by single molecule photobleaching

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Nanometric molecular separation measurements by single molecule photobleaching

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