Visualizing Protein-Protein Interactions by Non-Fitting and Easy FRET-FLIM Approaches

On-demand webinar by Dr. Sergi Padilla-Parra & Dr. Zhongxiang Jiang

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Understanding molecular interactions in living cells is crucial for deciphering molecular mechanisms underlying most cellular functions. The gold standard for studying protein-protein interaction is Förster resonance energy transfer (FRET). Although there are several approaches to demonstrate FRET in biological samples, using fluorescence lifetime imaging microscopy (FLIM) allows for a straightforward quantification of FRET based on the behavior of donor-only fluorescence.

About the Webinar

In this Webinar, Dr. Padilla-Parra shows how much information you can harness from FRET. He explains how lifetime-based non-fitting approaches such as minimal fraction of interacting donors (mFD) can provide a direct readout of protein-protein interactions in the cellular environment over time. He also talks about some of the limitations of classical FRET approaches.

After the presentation, Dr. Zhongxiang Jiang gives a live showcase at the STELLARIS 5 confocal system with an integrated White Light Laser (WLL), combined with the proprietary Acousto-optical Beam Splitter (AOBS) and new Power HyD S detectors. He shows the straightforward application of lifetime-based non-fitting FRET using the new and unique TauSense technology.

What to expect in the webinar

Key learnings

  • how to investigate protein-protein interaction in live samples;
  • ways to get the best out of FRET using fluorescence lifetime information;
  • how non-fitting approaches can be powerful tools for functional imaging;
  • what the minimal fraction of interacting donors (mFD) can tell you about molecular interactions in cells;
  • practical insights into using the STELLARIS 5 confocal system.

The Speakers

Dr. Sergi Padilla-Parra

After his MSc (major: Chemistry) obtained in Lund University (Sweden) he moved to the Neils Bohr Institute (Copenhagen, Denmark) to apply mathematical modelling to biological systems (from signaling to actin polymerization: towards a simplified mathematical model).

After that, he enrolled in a Research and Training Network on the Marie Curie FP7 (Institut Jacques Monod, Paris, 2006–2009) to apply his expertise in mathematics developing new methods in microscopy applied to complex biological problems (Regulation of actin assembly). In order to gain experience in biology and apply cutting edge microscopy approaches to retrovirus entry he moved to Emory University, Atlanta (2010–2012). After a period of two years he moved back to Europe to the University of Rennes in France, to implement the use of FRET biosensors in living embryos. Since April 2013, he is a principal investigator and leader of the Cellular Imaging Platform (CIP) at the Welcome Trust Centre for Human Genetics (WTCHG), University of Oxford. He develops novel quantitative microscopy approaches and apply them to better understand how some enveloped viruses (e.g. VSV, HIV or IAV) enter the host cell.

Dr. Zhongxiang Jiang

Zhongxiang Jiang received his PhD in cell biology in Karlsruhe Institute of Technology (KIT). After graduation, he joined Leica Microsystems in 2011.

As application manager for confocal microscopy, he is experienced in general confocal microscopy, multiphoton microscopy, Coherent Raman Scattering (CRS) technique and the field of fluorescence lifetime imaging (FLIM) and fluorescence correlation spectroscopy (FCS).

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