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A 7 µm mini-beam improves diffraction data from small or imperfect crystals of macromolecules

A simple apparatus for achieving beam sizes in the range 5–­10 µm on a synchrotron beamline was implemented in combination with a small 125 × 25 µm focus. The resulting beam had sufficient flux for crystallographic data collection from samples smaller than 10 × 10 × 10 µm. Sample data were collected...

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Autores principales: Sanishvili, Ruslan, Nagarajan, Venugopalan, Yoder, Derek, Becker, Michael, Xu, Shenglan, Corcoran, Stephen, Akey, David L., Smith, Janet L., Fischetti, Robert F.
Formato: Texto
Lenguaje:English
Publicado: International Union of Crystallography 2008
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2631116/
https://www.ncbi.nlm.nih.gov/pubmed/18391409
http://dx.doi.org/10.1107/S0907444908001741
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author Sanishvili, Ruslan
Nagarajan, Venugopalan
Yoder, Derek
Becker, Michael
Xu, Shenglan
Corcoran, Stephen
Akey, David L.
Smith, Janet L.
Fischetti, Robert F.
author_facet Sanishvili, Ruslan
Nagarajan, Venugopalan
Yoder, Derek
Becker, Michael
Xu, Shenglan
Corcoran, Stephen
Akey, David L.
Smith, Janet L.
Fischetti, Robert F.
author_sort Sanishvili, Ruslan
collection PubMed
description A simple apparatus for achieving beam sizes in the range 5–­10 µm on a synchrotron beamline was implemented in combination with a small 125 × 25 µm focus. The resulting beam had sufficient flux for crystallographic data collection from samples smaller than 10 × 10 × 10 µm. Sample data were collected representing three different scenarios: (i) a complete 2.0 Å data set from a single strongly diffracting microcrystal, (ii) a complete and redundant 1.94 Å data set obtained by merging data from six microcrystals and (iii) a complete 2.24 Å data set from a needle-shaped crystal with less than 12 × 10 µm cross-section and average diffracting power. The resulting data were of high quality, leading to well refined structures with good electron-density maps. The signal-to-noise ratios for data collected from small crystals with the mini-beam were significantly higher than for equivalent data collected from the same crystal with a 125 × 25 µm beam. Relative to this large beam, use of the mini-beam also resulted in lower refined crystal mosaicities. The mini-beam proved to be advantageous for inhomogeneous large crystals, where better ordered regions could be selected by the smaller beam.
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spelling pubmed-26311162009-03-05 A 7 µm mini-beam improves diffraction data from small or imperfect crystals of macromolecules Sanishvili, Ruslan Nagarajan, Venugopalan Yoder, Derek Becker, Michael Xu, Shenglan Corcoran, Stephen Akey, David L. Smith, Janet L. Fischetti, Robert F. Acta Crystallogr D Biol Crystallogr Research Papers A simple apparatus for achieving beam sizes in the range 5–­10 µm on a synchrotron beamline was implemented in combination with a small 125 × 25 µm focus. The resulting beam had sufficient flux for crystallographic data collection from samples smaller than 10 × 10 × 10 µm. Sample data were collected representing three different scenarios: (i) a complete 2.0 Å data set from a single strongly diffracting microcrystal, (ii) a complete and redundant 1.94 Å data set obtained by merging data from six microcrystals and (iii) a complete 2.24 Å data set from a needle-shaped crystal with less than 12 × 10 µm cross-section and average diffracting power. The resulting data were of high quality, leading to well refined structures with good electron-density maps. The signal-to-noise ratios for data collected from small crystals with the mini-beam were significantly higher than for equivalent data collected from the same crystal with a 125 × 25 µm beam. Relative to this large beam, use of the mini-beam also resulted in lower refined crystal mosaicities. The mini-beam proved to be advantageous for inhomogeneous large crystals, where better ordered regions could be selected by the smaller beam. International Union of Crystallography 2008-04-01 2008-03-19 /pmc/articles/PMC2631116/ /pubmed/18391409 http://dx.doi.org/10.1107/S0907444908001741 Text en © International Union of Crystallography 2008
spellingShingle Research Papers
Sanishvili, Ruslan
Nagarajan, Venugopalan
Yoder, Derek
Becker, Michael
Xu, Shenglan
Corcoran, Stephen
Akey, David L.
Smith, Janet L.
Fischetti, Robert F.
A 7 µm mini-beam improves diffraction data from small or imperfect crystals of macromolecules
title A 7 µm mini-beam improves diffraction data from small or imperfect crystals of macromolecules
title_full A 7 µm mini-beam improves diffraction data from small or imperfect crystals of macromolecules
title_fullStr A 7 µm mini-beam improves diffraction data from small or imperfect crystals of macromolecules
title_full_unstemmed A 7 µm mini-beam improves diffraction data from small or imperfect crystals of macromolecules
title_short A 7 µm mini-beam improves diffraction data from small or imperfect crystals of macromolecules
title_sort 7 µm mini-beam improves diffraction data from small or imperfect crystals of macromolecules
topic Research Papers
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2631116/
https://www.ncbi.nlm.nih.gov/pubmed/18391409
http://dx.doi.org/10.1107/S0907444908001741
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