Adaptations of the next generation of super-resolution microscopy: lessons learnt from imaging intracellular signalling nanodomains.
This talk outlines a decade of research which has utilised s
ingle molecule localisation microscopy to study intracellular ‘nanodomains’ which form the structural units of calcium
signalling seen in muscle cells. Adaptation of chemical photo-switching of aromatic fluorescent labels in the early experiments allowed me and my colleagues to produce paradigm-shifting super-resolution images of nanodomains of the heart over a decade ago. However, over the last few years, there has been a wave of newer super-resolution imaging methods which have transcended the level of spatial detail (resolution) in imaging nanodomains.
We are now able to map the precise positions of the ion channels and regulatory proteins, quantify protein co-clustering at single-protein precision, and identify their individual biochemical signatures.
This talk will detail the adaptation of DNA-PAINT, Enhanced Expansion Microscopy and sandSTORM, techniques which exploit the chemical and material properties of DNA, acrylamide-base hydrogels and fluorescent nano-diamonds in recent experiments which have discovered molecular-scale remodelling of nanodomains in pathologies such as right heart failure.