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MICROSCOPY & IMAGING


The new platform for 3D super-resolution microscopy from Zeiss


with other members of the team at ETHZ, Pinotsi has successfully deployed a variety of innovative technologies to further the study of Parkinson’s and Alzheimer’s diseases – and she continues working in the fields of super-resolution microscopy and spectroscopy for physical, chemical and life science applications. “Tese techniques have broken the limits of conventional microscopy imposed by the diffraction of light and allow us to ‘see’ molecular mechanisms and biological processes with nanometre resolution, even in live specimens,” she says. “One such technique is STORM – with which we can achieve unprecedented


spatial resolution, for a light microscopy technique, down to 20-30nm,” she adds.


NUCLEAR MEMBRANE Another of the most interesting recent developments in this field is at the University of Pittsburgh, where a team of researchers has employed STORM microscopy to investigate the structure of the nuclear envelope – which is made up of the inner and outer nuclear membrane and a protein meshwork, known as the nuclear lamina, that lies just beneath the inner nuclear membrane. As Dr Quasar Padiath, associate


professor in the Department of Human The researchers in Pittsburg who have employed STORM 54 www.scientistlive.com


Genetics at Pitt Graduate School of Public Health, explains, the nuclear lamina is composed of distinct proteins called lamins and is found in all metazoan cells. In addition to playing a critical role in regulating the shape and structural integrity of the nuclear envelope, it also has important functions in regulating gene expression through chromatin interactions and integrating cytoskeletal dynamics within the cell. In vertebrates, there are two major classes of lamins, the A and the B type. While previous research has suggested that the A and B type lamins form independent networks, whether they exhibit any distinct spatial organisation or were randomly distributed was unknown. “We attempted to elucidate the organisation of the distinct lamin proteins using STORM microscopy, which provides a significantly higher resolution than conventional light microscopy, while allowing for the simultaneous identification of multiple molecular species that are possible with other high-resolution imaging techniques such as electron microscopy (EM),” says Padiath. “STORM is based on single molecule localisation approach. In this work, we used both a custom-built STORM system and a commercially available STORM system from Nikon (N-STORM). Tis approach relies on turning ‘on’ a small subset of densely labelled fluorescent emitters, followed by precise localisation of their central positions at the precision


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