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Additional Ultrathin Carbon Layer

An ultrathin (2-3 nm) carbon layer applied to grids is widely recommended to improve particle distribution1,2,3 and has been shown to improve data quality when applied to both QUANTIFOIL® 4 Holey Carbon Films and UltrAuFoil® 5 Gold Supports. Shown on the right is the structure of Photorhabdus luminescens TcdA1, a large multisubunit complex toxin, with activity against insects, whose structure was determined using images collected on QUANTIFOIL® R 2/1 grids with a 2 nm additional carbon layer6.

Many biomolecules have a strong affinity for carbon, and a number of factors contribute to the improved specimen quality:

  • Increased numbers of particles due to adsorption of biomolecules onto carbon prior to blotting.
  • Improved particle distribution due to interaction with carbon surface across the hole.
  • Reduction in the number of particles adopting a preferred orientation.

An additional carbon layer can be added to any QUANTIFOIL® holey carbon film support on request. The most popular thickness is 2 nm, but thicker, more stable, 3 and 5 nm layers are also available, and even 10 nm is possible. If you are interested in adding an ultrathin carbon layer to UltrAuFoil® gold supports, please contact us for further details.  

References

  1. Passmore, LA and Russo, CJ. Methods Enzymol. 579: 51-86 (2016).
  2. Drulyte, I et alActa Cryst D74: 560-571 (2018).
  3. Thompson, RF et alMethods 100: 3-15 (2016).
  4. Rawson, S et alStructure 23: 461-471 (2015).
  5. Russo, CJ and Passmore, LA. J. Struct. Biol. 193:33-44 (2016).
  6. Leidreiter, F et alSci. Adv. 5: eaax6497 (2019).

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