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Capture and X-ray diffraction studies of protein microcrystals in a microfluidic trap array
X-ray free-electron lasers (XFELs) promise to enable the collection of interpretable diffraction data from samples that are refractory to data collection at synchrotron sources. At present, however, more efficient sample-delivery methods that minimize the consumption of microcrystalline material are...
Autores principales: | , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
International Union of Crystallography
2015
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4388268/ https://www.ncbi.nlm.nih.gov/pubmed/25849403 http://dx.doi.org/10.1107/S1399004715002308 |
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author | Lyubimov, Artem Y. Murray, Thomas D. Koehl, Antoine Araci, Ismail Emre Uervirojnangkoorn, Monarin Zeldin, Oliver B. Cohen, Aina E. Soltis, S. Michael Baxter, Elizabeth L. Brewster, Aaron S. Sauter, Nicholas K. Brunger, Axel T. Berger, James M. |
author_facet | Lyubimov, Artem Y. Murray, Thomas D. Koehl, Antoine Araci, Ismail Emre Uervirojnangkoorn, Monarin Zeldin, Oliver B. Cohen, Aina E. Soltis, S. Michael Baxter, Elizabeth L. Brewster, Aaron S. Sauter, Nicholas K. Brunger, Axel T. Berger, James M. |
author_sort | Lyubimov, Artem Y. |
collection | PubMed |
description | X-ray free-electron lasers (XFELs) promise to enable the collection of interpretable diffraction data from samples that are refractory to data collection at synchrotron sources. At present, however, more efficient sample-delivery methods that minimize the consumption of microcrystalline material are needed to allow the application of XFEL sources to a wide range of challenging structural targets of biological importance. Here, a microfluidic chip is presented in which microcrystals can be captured at fixed, addressable points in a trap array from a small volume (<10 µl) of a pre-existing slurry grown off-chip. The device can be mounted on a standard goniostat for conducting diffraction experiments at room temperature without the need for flash-cooling. Proof-of-principle tests with a model system (hen egg-white lysozyme) demonstrated the high efficiency of the microfluidic approach for crystal harvesting, permitting the collection of sufficient data from only 265 single-crystal still images to permit determination and refinement of the structure of the protein. This work shows that microfluidic capture devices can be readily used to facilitate data collection from protein microcrystals grown in traditional laboratory formats, enabling analysis when cryopreservation is problematic or when only small numbers of crystals are available. Such microfluidic capture devices may also be useful for data collection at synchrotron sources. |
format | Online Article Text |
id | pubmed-4388268 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | International Union of Crystallography |
record_format | MEDLINE/PubMed |
spelling | pubmed-43882682015-05-06 Capture and X-ray diffraction studies of protein microcrystals in a microfluidic trap array Lyubimov, Artem Y. Murray, Thomas D. Koehl, Antoine Araci, Ismail Emre Uervirojnangkoorn, Monarin Zeldin, Oliver B. Cohen, Aina E. Soltis, S. Michael Baxter, Elizabeth L. Brewster, Aaron S. Sauter, Nicholas K. Brunger, Axel T. Berger, James M. Acta Crystallogr D Biol Crystallogr Research Papers X-ray free-electron lasers (XFELs) promise to enable the collection of interpretable diffraction data from samples that are refractory to data collection at synchrotron sources. At present, however, more efficient sample-delivery methods that minimize the consumption of microcrystalline material are needed to allow the application of XFEL sources to a wide range of challenging structural targets of biological importance. Here, a microfluidic chip is presented in which microcrystals can be captured at fixed, addressable points in a trap array from a small volume (<10 µl) of a pre-existing slurry grown off-chip. The device can be mounted on a standard goniostat for conducting diffraction experiments at room temperature without the need for flash-cooling. Proof-of-principle tests with a model system (hen egg-white lysozyme) demonstrated the high efficiency of the microfluidic approach for crystal harvesting, permitting the collection of sufficient data from only 265 single-crystal still images to permit determination and refinement of the structure of the protein. This work shows that microfluidic capture devices can be readily used to facilitate data collection from protein microcrystals grown in traditional laboratory formats, enabling analysis when cryopreservation is problematic or when only small numbers of crystals are available. Such microfluidic capture devices may also be useful for data collection at synchrotron sources. International Union of Crystallography 2015-03-27 /pmc/articles/PMC4388268/ /pubmed/25849403 http://dx.doi.org/10.1107/S1399004715002308 Text en © Lyubimov et al. 2015 http://creativecommons.org/licenses/by/2.0/uk/ This is an open-access article distributed under the terms of the Creative Commons Attribution Licence, which permits unrestricted use, distribution, and reproduction in any medium, provided the original authors and source are cited. |
spellingShingle | Research Papers Lyubimov, Artem Y. Murray, Thomas D. Koehl, Antoine Araci, Ismail Emre Uervirojnangkoorn, Monarin Zeldin, Oliver B. Cohen, Aina E. Soltis, S. Michael Baxter, Elizabeth L. Brewster, Aaron S. Sauter, Nicholas K. Brunger, Axel T. Berger, James M. Capture and X-ray diffraction studies of protein microcrystals in a microfluidic trap array |
title | Capture and X-ray diffraction studies of protein microcrystals in a microfluidic trap array |
title_full | Capture and X-ray diffraction studies of protein microcrystals in a microfluidic trap array |
title_fullStr | Capture and X-ray diffraction studies of protein microcrystals in a microfluidic trap array |
title_full_unstemmed | Capture and X-ray diffraction studies of protein microcrystals in a microfluidic trap array |
title_short | Capture and X-ray diffraction studies of protein microcrystals in a microfluidic trap array |
title_sort | capture and x-ray diffraction studies of protein microcrystals in a microfluidic trap array |
topic | Research Papers |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4388268/ https://www.ncbi.nlm.nih.gov/pubmed/25849403 http://dx.doi.org/10.1107/S1399004715002308 |
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