Room-temperature macromolecular crystallography using a micro-patterned silicon chip with minimal background scattering

Recent success at X‐ray free‐electron lasers has led to serial crystallography experiments staging a comeback at synchrotron sources as well. With crystal lifetimes typically in the millisecond range and the latest‐generation detector technologies with high framing rates up to 1 kHz, fast sample exc...

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Vydané v:	Journal of applied crystallography Ročník 49; číslo 3; s. 968 - 975
Hlavní autori:	Roedig, Philip, Duman, Ramona, Sanchez-Weatherby, Juan, Vartiainen, Ismo, Burkhardt, Anja, Warmer, Martin, David, Christian, Wagner, Armin, Meents, Alke
Médium:	Journal Article
Jazyk:	English
Vydavateľské údaje:	5 Abbey Square, Chester, Cheshire CH1 2HU, England International Union of Crystallography 01.06.2016
Predmet:	crystallography on a chip Research Papers room-temperature crystallography synchrotron serial crystallography X-ray radiation damage crystallography on a chip room-temperature crystallography X-ray radiation damage synchrotron serial crystallography
ISSN:	1600-5767, 0021-8898, 1600-5767
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Shrnutí:	Recent success at X‐ray free‐electron lasers has led to serial crystallography experiments staging a comeback at synchrotron sources as well. With crystal lifetimes typically in the millisecond range and the latest‐generation detector technologies with high framing rates up to 1 kHz, fast sample exchange has become the bottleneck for such experiments. A micro‐patterned chip has been developed from single‐crystalline silicon, which acts as a sample holder for up to several thousand microcrystals at a very low background level. The crystals can be easily loaded onto the chip and excess mother liquor can be efficiently removed. Dehydration of the crystals is prevented by keeping them in a stream of humidified air during data collection. Further sealing of the sample holder, for example with Kapton, is not required. Room‐temperature data collection from insulin crystals loaded onto the chip proves the applicability of the chip for macromolecular crystallography. Subsequent structure refinements reveal no radiation‐damage‐induced structural changes for insulin crystals up to a dose of 565.6 kGy, even though the total diffraction power of the crystals has on average decreased to 19.1% of its initial value for the same dose. A decay of the diffracting power by half is observed for a dose of D1/2 = 147.5 ± 19.1 kGy, which is about 1/300 of the dose before crystals show a similar decay at cryogenic temperatures. A micro‐patterned sample holder of single‐crystalline silicon, loaded with multiple protein crystals which are exposed to a humidified gas stream, allows high‐quality room‐temperature data collection.
Bibliografia:	istex:1356AC163AEAB1CE3A3CF6A5773E647E1350FC6A ark:/67375/WNG-RXW82MKF-1 ArticleID:JCR2FS5133 ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	1600-5767 0021-8898 1600-5767
DOI:	10.1107/S1600576716006348