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@ARTICLE{Gicquel:402484,
author = {Gicquel, Yannig and Schubert, Robin and Kapis, Svetlana and
Bourenkov, Gleb and Schneider, Thomas and Perbandt, Markus
and Betzel, Christian and Chapman, Henry N. and Heymann,
Michael},
title = {{M}icrofluidic {C}hips for {I}n {S}itu {C}rystal {X}-ray
{D}iffraction and {I}n {S}itu {D}ynamic {L}ight {S}cattering
for {S}erial {C}rystallography},
journal = {Journal of visualized experiments},
volume = {134},
issn = {1940-087X},
address = {[S.l.]},
reportid = {PUBDB-2018-01851},
pages = {e57133},
year = {2018},
note = {PIF-2015-46},
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 fixed target serial X-ray crystallography at room
temperature. The protocol provides custom scripts to process
diffraction datasets using a suite of software tools to
solve and refine the protein crystal structure. This
approach avoids the artefacts possibly induced during
cryo-preservation or manual crystal handling in conventional
crystallography experiments. We present and compare three
protein structures that were solved using small crystals
with dimensions of approximately 10-20 μm grown in chip. By
crystallizing and diffracting in situ, handling and hence
mechanical disturbancesof fragile crystals is minimized. The
protocol details how to fabricate a custom X-ray transparent
microfluidic chip suitable for in situ serial
crystallography. As almost every crystal can be used for
diffraction data collection, these microfluidic chips are a
very efficient crystal delivery method.},
cin = {CFEL-I / UNI/CUI / EMBL},
ddc = {570},
cid = {I:(DE-H253)CFEL-I-20161114 / $I:(DE-H253)UNI_CUI-20121230$
/ I:(DE-H253)EMBL-20120731},
pnm = {6215 - Soft Matter, Health and Life Sciences (POF3-621) /
6G3 - PETRA III (POF3-622) / DFG project 194651731 - EXC
1074: Hamburger Zentrum für ultraschnelle Beobachtung
(CUI): Struktur, Dynamik und Kontrolle von Materie auf
atomarer Skala (194651731) / 05K13GU7 - Test eines "Serial
Femtosecond Crystallography (SFX)"-Messtandes am
Europäischen Elektronenlaser XFEL (BMBF-05K13GU7)},
pid = {G:(DE-HGF)POF3-6215 / G:(DE-HGF)POF3-6G3 /
G:(GEPRIS)194651731 / G:(DE-H253)BMBF-05K13GU7},
experiment = {EXP:(DE-H253)CFEL-Exp-20150101 /
EXP:(DE-H253)P-P14-20150101},
typ = {PUB:(DE-HGF)16},
pubmed = {pmid:29757285},
UT = {WOS:000444051300047},
doi = {10.3791/57133},
url = {https://bib-pubdb1.desy.de/record/402484},
}