Live cell super resolution imaging with the IISc, Bangalore

Deepak Nair together with Muthukumaran Venkatachalapathy of the Indian Institute of Science, Bangalore, have just reported the use of The Twinkle Factory‘s FAST for super resolution imaging in live cell.

See in Nanoscale: Live cell super resolution imaging by radial fluctuations using fluorogen binding tags. Nanoscale, 2019, 11, 3626-3632.

Abstract

Fluorescence-Activating and absorption-Shifting Tag (FAST) is a novel genetically encoded optical highlighter probe. Since the fluorescence of FAST originates from the stochastic and reversible diffusive association of a fluorogenic ligand, we investigate the application of FAST using Super-Resolution Radial Fluctuations (SRRF) to achieve routine imaging below the diffraction limit in a widefield epifluorescence microscope. We show that intensity fluctuation analysis like SRRF allows the imaging of FAST-tagged proteins with sub – 100 nm resolution in live cells. FAST co-labeled with conventional fluorophores enables real time multicolour 2D and 3D super-resolution imaging, indicating that FAST can be used for the observation of sub-diffraction limited structures in both living and fixed samples

Continue reading → Live cell super resolution imaging with the IISc, Bangalore

Want to image membrane proteins… without imaging?

Arnaud Gautier has just published how The Twinkle Factory may help to differentially label cell-surface and intracellular membrane protein based on the chemical-genetic fluorescent marker FAST. This approach allows the study of protein trafficking at the plasma membrane with various fluorometric techniques, including flow cytometry, with no imaging!

Check Bioconjugate Chem.: Fluorogenic Probing of Membrane Protein Trafficking. Bioconjug. Chem. (2018) 29(6), 1823-1828

Abstract

Methods to differentially label cell-surface and intracellular membrane proteins are indispensable for understanding their function and the regulation of their trafficking. We present an efficient strategy for the rapid and selective fluorescent labeling of membrane proteins based on the chemical-genetic fluorescent marker FAST (fluorescence-activating and absorption-shifting tag). Cell-surface FAST-tagged proteins could be selectively and rapidly labeled using fluorogenic membrane-impermeant 4-hydroxybenzylidene rhodanine (HBR) analogs. This approach allows the study of protein trafficking at the plasma membrane with various fluorometric techniques, and opens exciting prospects for the high-throughput screening of small molecules able to restore disease-related trafficking defects.