Micropipette aspiration as a tool for single-particle X-ray imaging and diffraction

J Synchrotron Radiat. 2023 Jul 1;30(Pt 4):788-795. doi: 10.1107/S1600577523003685. Epub 2023 May 26.

Abstract

A sample environment and manipulation tool is presented for single-particle X-ray experiments in an aqueous environment. The system is based on a single water droplet, positioned on a substrate that is structured by a hydrophobic and hydrophilic pattern to stabilize the droplet position. The substrate can support several droplets at a time. Evaporation is prevented by covering the droplet by a thin film of mineral oil. In this windowless fluid which minimizes background signal, single particles can be probed and manipulated by micropipettes, which can easily be inserted and steered in the droplet. Holographic X-ray imaging is shown to be well suited to observe and monitor the pipettes, as well as the droplet surface and the particles. Aspiration and force generation are also enabled based on an application of controlled pressure differences. Experimental challenges are addressed and first results are presented, obtained at two different undulator endstations with nano-focused beams. Finally, the sample environment is discussed in view of future coherent imaging and diffraction experiments with synchrotron radiation and single X-ray free-electron laser pulses.

Keywords: holographic X-ray imaging; micropipette aspiration; sample delivery of biophysical samples; single-particle diffraction.

MeSH terms

  • Holography*
  • Lasers*
  • Radiography
  • Synchrotrons
  • Water / chemistry
  • X-Ray Diffraction
  • X-Rays

Substances

  • Water