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. 2016 May 30;24(11):11515-30.
doi: 10.1364/OE.24.011515.

Three-dimensional-printed gas dynamic virtual nozzles for x-ray laser sample delivery

Garrett Nelson, Richard A Kirian, Uwe Weierstall, Nadia A Zatsepin, Tomáš Faragó, Tilo Baumbach, Fabian Wilde, Fabian B P Niesler, Benjamin Zimmer, Izumi Ishigami, Masahide Hikita, Saša Bajt, Syun-Ru Yeh, Denis L Rousseau, Henry N Chapman, John C H Spence, Michael Heymann

Three-dimensional-printed gas dynamic virtual nozzles for x-ray laser sample delivery

Garrett Nelson et al. Opt Express. .

Abstract

Reliable sample delivery is essential to biological imaging using X-ray Free Electron Lasers (XFELs). Continuous injection using the Gas Dynamic Virtual Nozzle (GDVN) has proven valuable, particularly for time-resolved studies. However, many important aspects of GDVN functionality have yet to be thoroughly understood and/or refined due to fabrication limitations. We report the application of 2-photon polymerization as a form of high-resolution 3D printing to fabricate high-fidelity GDVNs with submicron resolution. This technique allows rapid prototyping of a wide range of different types of nozzles from standard CAD drawings and optimization of crucial dimensions for optimal performance. Three nozzles were tested with pure water to determine general nozzle performance and reproducibility, with nearly reproducible off-axis jetting being the result. X-ray tomography and index matching were successfully used to evaluate the interior nozzle structures and identify the cause of off-axis jetting. Subsequent refinements to fabrication resulted in straight jetting. A performance test of printed nozzles at an XFEL provided high quality femtosecond diffraction patterns.

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Figures

Fig. 1
Fig. 1
(a) Bright field microscopy image from a traditional GDVN fabricated from flame polished glass capillaries. (b) Inset stroboscopic image of triggered droplets. (c) Image of nozzle components when immersed into an index-matched medium (glycerol), showing an undistorted view of the gas aperture profile and inner capillary position, as well as a laser-cut Kapton spacer for centering. The end of the outer glass capillary has been ground to a bevel to allow unshadowed wide angle X-ray diffraction. The contour indicated by the red line was used to derive the CAD model for the 3D printed nozzle.
Fig. 2
Fig. 2
(a) CAD renderings of nozzle design. (b) Cross section diagram of the printed nozzle tip assembly with both inner and outer capillaries glued into place.
Fig. 3
Fig. 3
(a) IP-S resist 3D printed GDVN. Stitching interfaces between the 400x400x10 µm3 units are visible as vertical (arrow 1) and horizontal (arrow 2) slices through the device. Misalignment appears at the perimeter as magnified in the inset, presumably due to free-floating regions with no support underneath. (b) Index-matched image (nozzle immersed in glycerol) showing projection of both vertical (arrow 1) and horizontal (arrow 2) stitching lines, corresponding to those shown in (a). Horizontal and vertical stitching lines cause very little disruption to continuity of features on the inner sidewall (insert). (c) 3D printed GVDN next to a U.S. dime (arrow 3).
Fig. 4
Fig. 4
Imaging of 3D printed nozzle tips to evaluate printing quality. (a) Bright field microscopy image of a fully assembled nozzle submerged in glycerol for index matching. A thin resin deposition is visible at the gas-focusing orifice. Also visible is the snug fit between the polished glass sample supply capillary and the 3D printed nozzle. (b) Nozzle tip submerged in glycerol with air bubble trapped inside. The original edge of the sidewall profile is clearly distinguishable from the deposition and matches the sidewall profile from the original CAD contour as highlighted by the white outline (c). (d) X-ray transmission image of a dried 3D printed nozzle after development. The black line at the bottom results from X-ray reflection of the glass substrate on which the nozzle was printed. (e) Cross sectional views from 3D surface rendering from an X-ray tomography reconstruction. The cutting plane of the cross section in each image is tilted with respect to the nozzle flow axis to illustrate the uneven resin accumulation near the tip in detail. (f, g) 3D surface rendering of nozzle after development with revised protocol (three development-wash-rinse cycles as opposed to just one). The nozzle was completely cleared of residual resin.
Fig. 5
Fig. 5
(a) Photograph of mounting stage. (b) Diagram of mounting stage indicating the electronic micromanipulator with syringe needle applicator, vacuum tweezers apparatus, fitting for holding the sample capillary securely, and manual micromanipulator for positioning and inserting capillary into nozzle. (c) The 3D printed nozzle is held in place with the vacuum tweezers while the sample capillary is inserted. With the nozzle secured by the capillary the vacuum tweezers can be turned off, and the capillary is then glued in place by applying epoxy with an applicator that is connected to the electronic micromanipulator. (d) The 3D printed nozzle with the attached capillary is glued into a steeply beveled stainless steel tube.
Fig. 6
Fig. 6
Nozzle performance when jetting water in vacuum, comparing the use of one development-wash-rinse to the use of three cycles. An angular deviation of about 6° relative to the nozzle axis was observed when using one cycle, whereas when 3 cycles were used the measured angular deviation was less than 1°. Nozzles are rotated by 60° between subsequent images as indicated by the red line in the schematics on the left column.
Fig. 7
Fig. 7
(a) X-ray scattering from 3D printed nozzle with (blue) and without sample (green) compared to scattering from a glass capillary GDVN running the same sample (red). The mean radial intensities are plotted as solid lines. The shaded region is bounded by the mean absolute deviation from the mean. The broad peak centered on 3.3Å is from the aqueous buffer, and the small peak at ~5 Å is from the printed nozzle. (b) Background scattering from 3D printed nozzle without sample running, from 2 orientations of the nozzle, rotated by 30° about the nozzle axis.
Fig. 8
Fig. 8
Example of membrane protein (cytochrome c oxidase) crystal diffraction obtained using a 3D printed nozzle, with sharp Bragg spots extending to 4 Å. No difference was observed between the quality of diffraction from samples run in the printed nozzle compared to a glass capillary GDVN.
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