QBI Neuroscience Seminar: Quantitative super-resolution microscopy: how well can we count single molecules?
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- Dr Nela Durisic of the Queensland Brain Institute, The University of Queensland hosts
- Quantitative super-resolution microscopy: how well can we count single molecules?
Abstract:
Light microscopy has undergone a revolution with the discovery of super-resolution imaging which has allowed biologists to study live cells at scales never observed before. These methods have resulted in vast and exciting new insights in the understanding the neuronal architecture including the structure of axons and neuronal synapses whose heterogeneous distribution of proteins is below the resolution limit of conventional microscopy. This is achieved through the use of photo-activated probes which can be “switched on” one at a time. Once structural information is revealed, the next significant step is to extract quantitative information on the number, size, distribution, and spatial organization of proteins. However, a thorough understanding of the complex photo-physical properties of these probes is needed for accurate quantification measurements. Here, I will explain how we can characterize the emission properties of GFP-like proteins which are used in single molecule localisation techniques such Photoactivated
Localisation Microscopy (PALM). Once a fluorescent probe has been characterized, it can then be used to measure properties such as protein stoichiometry, count protein numbers and determine spatial nano-organization of proteins in cells. In particular, I will focus on how these proteins can be used to count neurotransmitter receptors which are essential for the function and strength of the neuronal synapse.
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