Technologies

sandSTORM

This is a STORM protocol utilising nanometre-scale diamonds to gain unlimited imaging time in super-resolution microscopy. By-products of this adaptation have been a > 3-fold acceleration of the image acquisition, a >100-fold reduction in the light exposure to the sample and the departure from the need for unphysiological “fluorophore switching buffers” commonly used in STORM and similar techniques of localisation microscopy. See our paper on sandSTORM here.

 

‘Enhanced’ Expansion Microscopy

Expansion Microscopy (ExM) is super-resolution concept developed by Prof Ed Boyden and colleagues which allows a chemical imprint of cells and tissues onto a gel matrix. The gel matrix can then be inflated by a simple process of hydration to scale up the imprint, allowing its finer spatial features to be visualised with relatively simple fluorescence microscopy optics. We recently combined the “X10” ExM protocol, proposed by Sven Truckenbrodt and colleagues, with the resolution doubling of Airy Scan microscopy to image single ion channels in-situ. With a working in-plane resolution of ~ 15 nm and < 35 axially, this method allowed the reconstruction of three-dimensionally complex topologies of signalling nanodomains in cardiac cells. Read our latest paper on this protocol here.

DNA-PAINT

This is a single molecule localisation microscopy method which combines DNA oligo-nucleotide hybridisation with TIRF microscopy to achieve 5-15 nm in optical resolution for mapping protein, DNA and RNA targets. qPAINT is a target ‘counting’ algorithm which exploits the idea that DNA-PAINT localisation frequency, by the binding kinetics of ssDNA, is related to the underlying DNA target density. We have exploited this for ‘counting’ protein targets within intracellular nanodomains (paper in press with Cell Reports).

Read more about DNA-PAINT: Jungmann et al 2014, Auer et al 2017

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