MCFP Seminar: Introduction to Fluorescence Lifetime Imaging Microscopy (FLIM) and Fluorescence Correlation Spectroscopy (FCS).

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James Griffith

james.griffith@unimelb.edu.au

T: 0383440176

The Materials Characterisation and Fabrication Platform (MCFP) are currently upgrading their Nikon A1R+ Confocal Microscope to have Fluorescence Lifetime Imaging Microscopy (FLIM) and Fluorescence Correlation Spectroscopy (FCS) capabilities.

This seminar will act as an introduction to the techniques and how they can be used in your research.

Where: Chemical & Biomolecular Engineering Lecture Theatre, Building 165

Summary: Learn how FLIM and FCS can help you determine molecule interactions, interrogate the environment inside cellular organelles or measure concentration and size of particles in cells or materials.

Background: Typical fluorescence imaging relies on detection of photons to record intensity-based images. However, fluorescence lifetime, defined as the time the fluorophore remains in the excited state before returning to the ground state, opens up many more possibilities for analysis. The fluorescence lifetime is characteristic to each dye and its environment, providing the way for diverse techniques such as:

·         Local Environment Testing: Biological sensors such as pH, ion concentration and polarity

·         Detecting molecule interactions: Determine if molecules are interacting in living cells

·         Characterisation of materials: Use fluorescence lifetime to characterise materials

·         Multiplexing with multiple labels: FLIM allows discrimination of different fluorophores even when their fluorescence spectra overlaps

Fluorescence Correlation Spectroscopy (FCS) is a correlation analysis of temporal fluctuations of the fluorescence intensity. Biochemical parameters such as concentration and size or shape of the particle or viscosity of the environment can be determined.