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MICROSCOPY & IMAGING


in image-processing software have caused a ‘resolution revolution’, and a corresponding explosion in the number of researchers using cryo-EM.


WHAT DOES CRYO-ET HAVE TO OFFER? Cryo-ET goes a step further, offering 3D visualisations of molecular complexes in their native, fully hydrated environment, and is therefore ideally suited to reveal cellular organisation at molecular resolution. It’s like a CT scan for cells and biomolecules, using a series of 2D ‘slices’ through the sample to reconstruct a 3D image. Te holy grail of in situ structural biology is to obtain subnanometre or even near atomic resolution maps of entire cellular landscapes. Can this be achieved with cryo-ET? Wolfgang Baumeister researches cellular machines called macromolecular complexes at the Max Planck institute of Biochemistry in Munich. His team developed cryo-ET in the late 1980s, leading to the first successful mapping of macromolecular complexes in intact cells in 2000. For him, one of the main challenges in the uptake of cryo-ET is the start-up costs. He says, “To start a lab that can do cutting-edge research along the these lines, one has to make minimal investment of some something like €10 million. Tat’s affordable for larger institutions, but there are many other scientists out there who have interesting and challenging problems who cannot


make such an investment, and they need to be given access to it and to the technology.”


Te solution to the access problem is


to create national cryo-EM/ET facilities. In the UK, the Electron Bio-Imaging Centre (eBIC) was established at Diamond Light Source, the UK’s national synchrotron, with the award of a £15.6 million grant from the Wellcome Trust, the Medical Research Council (MRC) and the Biotechnology and Biological Sciences Research Council (BBSRC).


Dr Alistair Siebert docking an Autoloader Nanocab containing a 12 sample cassette


The Autoloader enables rapid sample exchange at liquid nitrogen temperature


Diamond and eBIC allow scientists to access state-of-the-art cryo-EM facilities, for both single particle analysis and cryo-tomography. Scientists are using onsite cryo-electron microscopes to study everything from complex virus structures to never-before-seen proteins.


Life Science director Dave Stuart outside Diamond Light Source


58 www.scientistlive.com


CRACKING THE HIV CODE Peijun Zhang is the director of eBIC, where her research focuses on both cryo-EM technology development and investigation of human pathogens such as HIV-1. According to UNAIDS, there were 36.9 million people living with HIV in 2017, of which 1.8 million were newly infected. Tere is currently no cure or vaccination for HIV, and 35.4 million have died from AIDS-related illnesses since the start of the epidemic. Te key to developing a cure for HIV lies in understanding its structure, and how it infects human cells, a complex code that has proved hard to crack.


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