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Dual-color metal-induced and förster resonance energy transfer for cell nanoscopy.

Molecular biology of the cell (2018-02-16)
Anna M Chizhik, Carina Wollnik, Daja Ruhlandt, Narain Karedla, Alexey I Chizhik, Lara Hauke, Dirk Hähnel, Ingo Gregor, Jörg Enderlein, Florian Rehfeldt
要旨

We report a novel method, dual color axial nanometric localization by Metal Induced Energy Transfer (dcMIET), and combine it with Förster Resonant Energy Transfer (FRET) for resolving structural details in cells on the molecular level. We demonstrate the capability of this method on cytoskeletal elements and adhesions in human mesenchymal stem cells (hMSCs). Our approach is based on Fluorescence-Lifetime-Imaging Microscopy (FLIM), and allows for precise determination of the 3D architecture of stress fibers anchoring at focal adhesions, thus yielding crucial information to understand cell-matrix mechanics. In addition to resolving nanometric structural details along the z-axis, we use FRET to gain precise information on the distance between actin and vinculin at focal adhesions.

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Sigma-Aldrich
抗-ウサギ-IgG - Atto 647N ヤギ宿主抗体, 1 mg/mL IgG
Sigma-Aldrich
Anti-Mouse IgG (whole molecule)−Atto 488 antibody produced in goat, ~1 mg/mL