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  • Simultaneous widefield single molecule orientation and FRET microscopy in cells.

Simultaneous widefield single molecule orientation and FRET microscopy in cells.

Optics express (2008-12-10)
S E D Webb, D J Rolfe, S R Needham, S K Roberts, D T Clarke, C I McLachlan, M P Hobson, M L Martin-Fernandez
ABSTRACT

We combine single molecule fluorescence orientation imaging with single-pair fluorescence resonance energy transfer microscopy, using a total internal reflection microscope. We show how angles and FRET efficiencies can be determined for membrane proteins at the single molecule level and provide data from the epidermal growth factor receptor system in cells.

MATERIALS
Product Number
Brand
Product Description

Supelco
Atto 647N maleimide, BioReagent, suitable for fluorescence, ≥90% (HPLC)
Sigma-Aldrich
Atto 647N azide, BioReagent, suitable for fluorescence
Sigma-Aldrich
Atto 647N amine, BioReagent, suitable for fluorescence
Sigma-Aldrich
Atto 647N-Biotin, BioReagent, suitable for fluorescence, ≥90.0% (HPLC)
Supelco
Atto 647N, BioReagent, suitable for fluorescence, ≥90.0% (HPLC)
Sigma-Aldrich
Atto 647N NHS ester, BioReagent, suitable for fluorescence, ≥90% (HPLC)