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	The high-throughput production of membrane proteins James Birch , Andrew Quigley Emerging Topics In Life Sciences 6 DOI: 10.1042/ETLS20210196 Open Access Show Abstract ▼ Abstract: Membrane proteins, found at the junctions between the outside world and the inner workings of the cell, play important roles in human disease and are used as biosensors. More than half of all therapeutics directly affect membrane protein function while nanopores enable DNA sequencing. The structural and functional characterisation of membrane proteins is therefore crucial. However, low levels of naturally abundant protein and the hydrophobic nature of membrane proteins makes production difficult. To maximise success, high-throughput strategies were developed that rely upon simple screens to identify successful constructs and rapidly exclude those unlikely to work. Parameters that affect production such as expression host, membrane protein origin, expression vector, fusion-tags, encapsulation reagent and solvent composition are screened in parallel. In this way, constructs with divergent requirements can be produced for a variety of structural applications. As structural techniques advance, sample requirements will change. Single-particle cryo-electron microscopy requires less protein than crystallography and as cryo-electron tomography and time-resolved serial crystallography are developed new sample production requirements will evolve. Here we discuss different methods used for the high-throughput production of membrane proteins for structural biology.	Oct 2021
B24-Cryo Soft X-ray Tomography	Cryo-soft X-ray tomography: using soft X-rays to explore the ultrastructure of whole cells Maria Harkiolaki , Michele C. Darrow , Matthew C. Spink , Ewelina Kosior , Kyle Dent , Elisabeth Duke Emerging Topics In Life Sciences DOI: 10.1042/ETLS20170086 Open Access Show Abstract ▼ Abstract: Cryo-soft X-ray tomography is an imaging technique that addresses the need for mesoscale imaging of cellular ultrastructure of relatively thick samples without the need for staining or chemical modification. It allows the imaging of cellular ultrastructure to a resolution of 25–40 nm and can be used in correlation with other imaging modalities, such as electron tomography and fluorescence microscopy, to further enhance the information content derived from biological samples. An overview of the technique, discussion of sample suitability and information about sample preparation, data collection and data analysis is presented here. Recent developments and future outlook are also discussed.	Mar 2018

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