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Self-Blotting Nanowire Grids for Cryo-EM Sample Preparation

  • Hui Wei (a1), Venkat Dandey (a1), Zhening Zhang (a1), Ashleigh Raczkowski (a1), Bridget Carragher (a1) and Clinton S. Potter (a1)...

Abstract

Almost every aspect of cryo electron microscopy (CryoEM) has been automated over the last few decades. One of the challenges that remains to be addressed is the robust and reliable preparation of vitrified specimens of suitable ice thickness. The development of a new self-blotting nanowire (Zhang et al., 2013) grid in conjunction with a piezo electric dispensing robot called Spotiton (Jain et al., 2012) enables spreading a sample to a thin film without the use of externally applied filter paper. This new approach has the advantage of using small amounts of protein material, resulting in large areas of ice of a well- defined thickness containing evenly distributed particles (Razinkov et al., 2016).

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References

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1. Zhang, F., Zhang, W.B., Shi, Z., Wang, D., Jin, J. & Jiang, L. (2013). Nanowire-haired inorganic membranes with superhydrophilicity and underwater ultralow adhesive superoleophobicity for high-efficiency oil/water separation. Adv. Mater 25, 41924198.
2. Jain, T., Sheehan, P., Crum, J., Carragher, B. & Potter, C.S. (2012). Spotiton: a prototype for an integrated inkjet dispense and vitrification system for cryo-TEM. J. Struct. Biol 179, 6875.
3. Razinkov, I, Dandey, VP, Wei, H, Zhang, Z, Melnekoff, D, Rice, WJ, Wigge, C, Potter, CS & Carragher, B (2016). A new method for vitrifying samples for cryoEM. J Struct Biol 195(2), 190198.
4. Russo, C.J. & Passmore, L.A. (2016). Ultrastable gold substrates: properties of a support for high-resolution electron cryomicroscopy of biological specimens. J. Struct. Biol. 193, 3344.

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