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A user-friendly goniometer-compatible fixed-target platform for macromolecular crystallography at synchrotrons

Ghosh, Swagatha ; Banacore, Analia ; Norder, Per ; Bjelčić, Monica LU ; Kabbinale, Arpitha ; Nileshwar, Padmini ; Wehlander, Gabrielle ; de Sanctis, Daniele ; Basu, Shibom and Orlans, Julien , et al. (2026) In Journal of Applied Crystallography 59. p.303-315
Abstract

Fixed-target platforms provide convenient support for microcrystals during serial X-ray crystallography studies using synchrotron radiation. Here, we describe a simple user-friendly 3D-printed support where the crystals are sandwiched between two layers of thin X-ray-transparent membrane resulting in very low scattering background. The platform is compatible with magnetic mounting onto the standard goniometer of macromolecular crystallography beamlines. Our design utilizes a 96-well frame that facilitates hanging-drop experiments directly on the membrane using conventional crystallization plates, thereby eliminating multiple pipetting and crystal handling steps. Crystals can be enclosed in a sandwich and packed into ‘cassettes’,... (More)

Fixed-target platforms provide convenient support for microcrystals during serial X-ray crystallography studies using synchrotron radiation. Here, we describe a simple user-friendly 3D-printed support where the crystals are sandwiched between two layers of thin X-ray-transparent membrane resulting in very low scattering background. The platform is compatible with magnetic mounting onto the standard goniometer of macromolecular crystallography beamlines. Our design utilizes a 96-well frame that facilitates hanging-drop experiments directly on the membrane using conventional crystallization plates, thereby eliminating multiple pipetting and crystal handling steps. Crystals can be enclosed in a sandwich and packed into ‘cassettes’, preventing the risk of the sample drying out during room-temperature transportation to synchrotron sources. The versatility of the platform is demonstrated by five structures solved using different crystallization and data-collection strategies. Lysozyme single-crystal rotational crystallography at room temperature is shown, as well as microcrystal serial data collection under cryogenic conditions. On-chip micro-crystallization is illustrated by use of a photosynthetic reaction center as an example. Finally, serial crystallography data collection at room temperature from microcrystals of the membrane protein cytochrome c oxidase crystallized in lipidic cubic phase is presented.

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organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
fixed-target chips, in situ crystallization, macromolecular crystallography, room-temperature protein structures, serial synchrotron X-ray crystallography
in
Journal of Applied Crystallography
volume
59
pages
13 pages
publisher
International Union of Crystallography
external identifiers
  • scopus:105037384770
  • pmid:41959872
ISSN
0021-8898
DOI
10.1107/S1600576725011513
language
English
LU publication?
yes
id
fd4b5080-0f58-4139-8c30-2d653ebafc25
date added to LUP
2026-05-25 12:01:47
date last changed
2026-06-08 12:56:08
@article{fd4b5080-0f58-4139-8c30-2d653ebafc25,
  abstract     = {{<p>Fixed-target platforms provide convenient support for microcrystals during serial X-ray crystallography studies using synchrotron radiation. Here, we describe a simple user-friendly 3D-printed support where the crystals are sandwiched between two layers of thin X-ray-transparent membrane resulting in very low scattering background. The platform is compatible with magnetic mounting onto the standard goniometer of macromolecular crystallography beamlines. Our design utilizes a 96-well frame that facilitates hanging-drop experiments directly on the membrane using conventional crystallization plates, thereby eliminating multiple pipetting and crystal handling steps. Crystals can be enclosed in a sandwich and packed into ‘cassettes’, preventing the risk of the sample drying out during room-temperature transportation to synchrotron sources. The versatility of the platform is demonstrated by five structures solved using different crystallization and data-collection strategies. Lysozyme single-crystal rotational crystallography at room temperature is shown, as well as microcrystal serial data collection under cryogenic conditions. On-chip micro-crystallization is illustrated by use of a photosynthetic reaction center as an example. Finally, serial crystallography data collection at room temperature from microcrystals of the membrane protein cytochrome c oxidase crystallized in lipidic cubic phase is presented.</p>}},
  author       = {{Ghosh, Swagatha and Banacore, Analia and Norder, Per and Bjelčić, Monica and Kabbinale, Arpitha and Nileshwar, Padmini and Wehlander, Gabrielle and de Sanctis, Daniele and Basu, Shibom and Orlans, Julien and Vallejos, Adams and Chavas, Leonard M.G. and Neutze, Richard and Brändén, Gisela}},
  issn         = {{0021-8898}},
  keywords     = {{fixed-target chips; in situ crystallization; macromolecular crystallography; room-temperature protein structures; serial synchrotron X-ray crystallography}},
  language     = {{eng}},
  pages        = {{303--315}},
  publisher    = {{International Union of Crystallography}},
  series       = {{Journal of Applied Crystallography}},
  title        = {{A user-friendly goniometer-compatible fixed-target platform for macromolecular crystallography at synchrotrons}},
  url          = {{http://dx.doi.org/10.1107/S1600576725011513}},
  doi          = {{10.1107/S1600576725011513}},
  volume       = {{59}},
  year         = {{2026}},
}