Microfluidic sorting of protein nanocrystals by size for X-ray free-electron laser diffraction

Bahige G. Abdallah, Nadia A. Zatsepin, Shatabdi Roy-Chowdhury, Jesse Coe, Chelsie E. Conrad, Katerina Dörner, Raymond G. Sierra, Hilary P. Stevenson, Fernanda Camacho-Alanis, Thomas D. Grant, Garrett Nelson, Daniel James, Guillermo Calero, Rebekka M. Wachter, John C H Spence, Uwe Weierstall, Petra Fromme, Alexandra Ros

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

The advent and application of the X-ray free-electron laser (XFEL) has uncovered the structures of proteins that could not previously be solved using traditional crystallography. While this new technology is powerful, optimization of the process is still needed to improve data quality and analysis efficiency. One area is sample heterogeneity, where variations in crystal size (among other factors) lead to the requirement of large data sets (and thus 10-100 mg of protein) for determining accurate structure factors. To decrease sample dispersity, we developed a high-throughput microfluidic sorter operating on the principle of dielectrophoresis, whereby polydisperse particles can be transported into various fluid streams for size fractionation. Using this microsorter, we isolated several milliliters of photosystem I nanocrystal fractions ranging from 200 to 600 nm in size as characterized by dynamic light scattering, nanoparticle tracking, and electron microscopy. Sorted nanocrystals were delivered in a liquid jet via the gas dynamic virtual nozzle into the path of the XFEL at the Linac Coherent Light Source. We obtained diffraction to ~4 Å resolution, indicating that the small crystals were not damaged by the sorting process. We also observed the shape transforms of photosystem I nanocrystals, demonstrating that our device can optimize data collection for the shape transform-based phasing method. Using simulations, we show that narrow crystal size distributions can significantly improve merged data quality in serial crystallography. From this proof-of-concept work, we expect that the automated size-sorting of protein crystals will become an important step for sample production by reducing the amount of protein needed for a high quality final structure and the development of novel phasing methods that exploit inter-Bragg reflection intensities or use variations in beam intensity for radiation damage-induced phasing. This method will also permit an analysis of the dependence of crystal quality on crystal size.

Original languageEnglish
Article number041719
JournalStructural Dynamics
Volume2
Issue number4
DOIs
Publication statusPublished - Jul 1 2015

Fingerprint

X ray lasers
Free electron lasers
classifying
Sorting
Microfluidics
free electron lasers
Nanocrystals
nanocrystals
Diffraction
proteins
Proteins
Crystals
diffraction
crystals
Photosystem I Protein Complex
x rays
Crystallography
crystallography
Gas dynamics
gas dynamics

ASJC Scopus subject areas

  • Condensed Matter Physics
  • Instrumentation
  • Radiation
  • Spectroscopy

Cite this

Abdallah, B. G., Zatsepin, N. A., Roy-Chowdhury, S., Coe, J., Conrad, C. E., Dörner, K., ... Ros, A. (2015). Microfluidic sorting of protein nanocrystals by size for X-ray free-electron laser diffraction. Structural Dynamics, 2(4), [041719]. https://doi.org/10.1063/1.4928688

Microfluidic sorting of protein nanocrystals by size for X-ray free-electron laser diffraction. / Abdallah, Bahige G.; Zatsepin, Nadia A.; Roy-Chowdhury, Shatabdi; Coe, Jesse; Conrad, Chelsie E.; Dörner, Katerina; Sierra, Raymond G.; Stevenson, Hilary P.; Camacho-Alanis, Fernanda; Grant, Thomas D.; Nelson, Garrett; James, Daniel; Calero, Guillermo; Wachter, Rebekka M.; Spence, John C H; Weierstall, Uwe; Fromme, Petra; Ros, Alexandra.

In: Structural Dynamics, Vol. 2, No. 4, 041719, 01.07.2015.

Research output: Contribution to journalArticle

Abdallah, BG, Zatsepin, NA, Roy-Chowdhury, S, Coe, J, Conrad, CE, Dörner, K, Sierra, RG, Stevenson, HP, Camacho-Alanis, F, Grant, TD, Nelson, G, James, D, Calero, G, Wachter, RM, Spence, JCH, Weierstall, U, Fromme, P & Ros, A 2015, 'Microfluidic sorting of protein nanocrystals by size for X-ray free-electron laser diffraction', Structural Dynamics, vol. 2, no. 4, 041719. https://doi.org/10.1063/1.4928688
Abdallah BG, Zatsepin NA, Roy-Chowdhury S, Coe J, Conrad CE, Dörner K et al. Microfluidic sorting of protein nanocrystals by size for X-ray free-electron laser diffraction. Structural Dynamics. 2015 Jul 1;2(4). 041719. https://doi.org/10.1063/1.4928688
Abdallah, Bahige G. ; Zatsepin, Nadia A. ; Roy-Chowdhury, Shatabdi ; Coe, Jesse ; Conrad, Chelsie E. ; Dörner, Katerina ; Sierra, Raymond G. ; Stevenson, Hilary P. ; Camacho-Alanis, Fernanda ; Grant, Thomas D. ; Nelson, Garrett ; James, Daniel ; Calero, Guillermo ; Wachter, Rebekka M. ; Spence, John C H ; Weierstall, Uwe ; Fromme, Petra ; Ros, Alexandra. / Microfluidic sorting of protein nanocrystals by size for X-ray free-electron laser diffraction. In: Structural Dynamics. 2015 ; Vol. 2, No. 4.
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