Microfluidic Chips for In Situ Crystal X-ray Diffraction and In Situ Dynamic Light Scattering for Serial Crystallography

Journal of Visualized Experiments : JoVE
Yannig GicquelMichael Heymann

Abstract

This protocol describes fabricating microfluidic devices with low X-ray background optimized for goniometer based fixed target serial crystallography. The devices are patterned from epoxy glue using soft lithography and are suitable for in situ X-ray diffraction experiments at room temperature. The sample wells are lidded on both sides with polymeric polyimide foil windows that allow diffraction data collection with low X-ray background. This fabrication method is undemanding and inexpensive. After the sourcing of a SU-8 master wafer, all fabrication can be completed outside of a cleanroom in a typical research lab environment. The chip design and fabrication protocol utilize capillary valving to microfluidically split an aqueous reaction into defined nanoliter sized droplets. This loading mechanism avoids the sample loss from channel dead-volume and can easily be performed manually without using pumps or other equipment for fluid actuation. We describe how isolated nanoliter sized drops of protein solution can be monitored in situ by dynamic light scattering to control protein crystal nucleation and growth. After suitable crystals are grown, complete X-ray diffraction datasets can be collected using goniometer based in situ fi...Continue Reading

Citations

Oct 17, 2019·The FEBS Journal·Feng-Zhu ZhaoDa-Chuan Yin
Aug 4, 2020·Acta Crystallographica. Section D, Structural Biology·Jose A GaviraChristoph Mueller-Dieckman
Oct 16, 2020·Lab on a Chip·Anne BremerMichael Heymann
Jul 19, 2019·Langmuir : the ACS Journal of Surfaces and Colloids·Mohammad VakiliMartin Trebbin
May 29, 2019·ACS Applied Materials & Interfaces·Hiromune EtoPetra Schwille

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Methods Mentioned

BETA
X-ray
chips
dynamic light scattering
chip
biopsy
light scattering

Software Mentioned

AutoCad
Origin9
XSCALE
ProtParam
Matlab
Refmac5
XPARM
Bash
XDS
MOLREP

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