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.
Molecular Biosensor and Imaging Center, Carnegie Mellon University, 4400 Fifth Avenue, Pittsburgh PA 15213.
James A J
JA
Fitzpatrick
Qi
Q
Yan
Jochen J
JJ
Sieber
Marcus
M
Dyba
Ulf
U
Schwarz
Chris
C
Szent-Gyorgyi
Carol A
CA
Woolford
Peter B
PB
Berget
Alan S
AS
Waggoner
Marcel P
MP
Bruchez