STED nanoscopy in living cells using Fluorogen Activating Proteins.
We demonstrate the effectiveness of a genetically encoded Malachite Green (MG) binding fluorogen activating protein (FAP) for live cell stimulated emission depletion nanoscopy (STED). Both extracellular and intracellular FAPs were tested in living cells using fluorogens with either membrane expressed FAP or as an intracellular FAP-actin fusion. Structures with FWHM of 110-122nm were observed. Depletion data however suggests a resolution of 70nm with the given instrument.
Fitzpatrick
James A J
JA
Molecular Biosensor and Imaging Center, Carnegie Mellon University, 4400 Fifth Avenue, Pittsburgh PA 15213.
Yan
Qi
Q
Sieber
Jochen J
JJ
Dyba
Marcus
M
Schwarz
Ulf
U
Szent-Gyorgyi
Chris
C
Woolford
Carol A
CA
Berget
Peter B
PB
Waggoner
Alan S
AS
Bruchez
Marcel P
MP