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Adam
Round
,
Pierre
Aller
,
Richard
Bean
,
Johan
Bielecki
,
Agata
Butryn
,
Nicholas E.
Devenish
,
Raphael
De Wijn
,
Thomas
Dietze
,
Katerina
Doerner
,
Fabio
Dall'Antonia
,
Gabriele
Giovanetti
,
Huijong
Han
,
Vincent
Hennicke
,
Chan
Kim
,
Yoonhee
Kim
,
Marco
Kloos
,
Jayanath C. P.
Koliyadu
,
Gabriel
Leen
,
Romain
Letrun
,
Luis
Lopez Morillo
,
Allen M.
Orville
,
Tim
Pakendorf
,
Marco
Ramilli
,
Nadja
Reimers
,
Patrick
Reinke
,
Juan
Sanchez-Weatherby
,
Tokushi
Sato
,
Robin
Schubert
,
Joachim
Schulz
,
Cedric
Signe Takem
,
Marcin
Sikorski
,
Prasad
Thute
,
Fabian
Trost
,
Oleksii
Turkot
,
Patrik
Vagovic
,
Mohammad
Vakili
,
Raul
Villanueva Guerrero
,
Henry N.
Chapman
,
Alke
Meents
,
Serguei
Molodtsov
,
Sakura
Pascarelli
,
Thomas
Tschentschera
,
Adrian
Mancuso
,
Pontus
Fischer
,
Sebastian
Guenther
Open Access
Abstract: The Single-Particle, Clusters and Biomolecules and Serial Femtosecond Crystallography (SPB/SFX) scientific instrument at the European X-Ray Free-Electron Laser (EuXFEL) became operational with user experiments in September 2017. The unique properties and capabilities of the EuXFEL, enabling megahertz data collection rates, provide more rapid data collection with improved statistics compared with other XFEL facilities. This improves the feasibility of obtaining multiple data points in time-resolved experiments and hence enables the observation of reactions in greater detail (molecular movies). In collaboration with the SFX User Consortium (SFX UC), the SPB/SFX instrument was designed to further increase user access and research outcomes. Focusing the pulses downstream of the first interaction region [described previously (Mancuso et al., 2019)], a second experiment plane is enabled, which allows for greater optimization and more efficient usage of available beam time. Additionally, the SFX UC provided further instrumentation to provide improved capabilities on SPB/SFX. The aim for additional and extended functionality for the second interaction region was to enable sample-efficient data collection at atmospheric pressure in an environment where the sample temperature and humidity can be controlled. This paper describes the extended capabilities of the downstream interaction region of the SPB/SFX instrument and its major components, in particular its X-ray focusing optics, vacuum to atmospheric pressure out-coupling, available sample delivery methods and 2D detector, and the supporting optical laser systems for pump–probe experiments.
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Nov 2025
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Zhou
Shen
,
Paul Lourdu
Xavier
,
Richard
Bean
,
Johan
Bielecki
,
Martin
Bergemann
,
Benedikt
Daurer
,
Tomas
Ekeberg
,
Armando D.
Estillore
,
Hans
Fangohr
,
Klaus
Giewekemeyer
,
Mikhail
Karnevskiy
,
Richard A.
Kirian
,
Henry
Kirkwood
,
Yoonhee
Kim
,
Jayanath C. P.
Koliyadu
,
Holger
Lange
,
Romain
Letrun
,
Jannik
Lübke
,
Abhishek
Mall
,
Thomas
Michelat
,
Andrew J.
Morgan
,
Nils
Roth
,
Amit K.
Samanta
,
Tokushi
Sato
,
Marcin
Sikorski
,
Florian
Schulz
,
Patrik
Vagovic
,
Tamme
Wollweber
,
Lena
Worbs
,
Filipe
Maia
,
Daniel A.
Horke
,
Jochen
Küpper
,
Adrian P.
Mancuso
,
Henry
Chapman
,
Kartik
Ayyer
,
N. Duane
Loh
Open Access
Abstract: Nanoparticles, exhibiting functionally relevant structural heterogeneity, are at the forefront of cutting-edge research. Now, high-throughput single-particle imaging (SPI) with X-ray free-electron lasers (XFELs) creates opportunities for recovering the shape distributions of millions of particles that exhibit functionally relevant structural heterogeneity. To realize this potential, three challenges have to be overcome: (1) simultaneous parametrization of structural variability in real and reciprocal spaces; (2) efficiently inferring the latent parameters of each SPI measurement; (3) scaling up comparisons between 105 structural models and 106 XFEL-SPI measurements. Here, we describe how we overcame these three challenges to resolve the nonequilibrium shape distributions within millions of gold nanoparticles imaged at the European XFEL. These shape distributions allowed us to quantify the degree of asymmetry in these particles, discover a relatively stable “shape envelope” among nanoparticles, discern finite-size effects related to shape-controlling surfactants, and extrapolate nanoparticles’ shapes to their idealized thermodynamic limit. Ultimately, these demonstrations show that XFEL SPI can help transform nanoparticle shape characterization from anecdotally interesting to statistically meaningful.
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May 2024
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Austin
Echelmeier
,
Jorvani
Cruz Villarreal
,
Marc
Messerschmidt
,
Daihyun
Kim
,
Jesse D.
Coe
,
Darren
Thifault
,
Sabine
Botha
,
Ana
Egatz-Gomez
,
Sahir
Gandhi
,
Gerrit
Brehm
,
Chelsie E.
Conrad
,
Debra T.
Hansen
,
Caleb
Madsen
,
Saša
Bajt
,
J. Domingo
Meza-Aguilar
,
Dominik
Oberthuer
,
Max O.
Wiedorn
,
Holger
Fleckenstein
,
Derek
Mendez
,
Juraj
Knoška
,
Jose M.
Martin-Garcia
,
Hao
Hu
,
Stella
Lisova
,
Aschkai
Allahgoli
,
Yaroslav
Gevorkov
,
Kartik
Ayyer
,
Steve
Aplin
,
Helen M.
Ginn
,
Heinz
Graafsma
,
Andrew J.
Morgan
,
Dominic
Greiffenberg
,
Alexander
Klujev
,
Torsten
Laurus
,
Jennifer
Poehlsen
,
Ulrich
Trunk
,
Davide
Mezza
,
Bernd
Schmitt
,
Manuela
Kuhn
,
Raimund
Fromme
,
Jolanta
Sztuk-Dambietz
,
Natascha
Raab
,
Steffen
Hauf
,
Alessandro
Silenzi
,
Thomas
Michelat
,
Chen
Xu
,
Cyril
Danilevski
,
Andrea
Parenti
,
Leonce
Mekinda
,
Britta
Weinhausen
,
Grant
Mills
,
Patrik
Vagovic
,
Yoonhee
Kim
,
Henry
Kirkwood
,
Richard
Bean
,
Johan
Bielecki
,
Stephan
Stern
,
Klaus
Giewekemeyer
,
Adam
Round
,
Joachim
Schulz
,
Katerina
Dörner
,
Thomas D.
Grant
,
Valerio
Mariani
,
Anton
Barty
,
Adrian P.
Mancuso
,
Uwe
Weierstall
,
John C. H.
Spence
,
Henry N.
Chapman
,
Nadia
Zatsepin
,
Petra
Fromme
,
Richard A.
Kirian
,
Alexandra
Ros
Open Access
Abstract: Serial femtosecond crystallography (SFX) with X-ray free electron lasers (XFELs) allows structure determination of membrane proteins and time-resolved crystallography. Common liquid sample delivery continuously jets the protein crystal suspension into the path of the XFEL, wasting a vast amount of sample due to the pulsed nature of all current XFEL sources. The European XFEL (EuXFEL) delivers femtosecond (fs) X-ray pulses in trains spaced 100 ms apart whereas pulses within trains are currently separated by 889 ns. Therefore, continuous sample delivery via fast jets wastes >99% of sample. Here, we introduce a microfluidic device delivering crystal laden droplets segmented with an immiscible oil reducing sample waste and demonstrate droplet injection at the EuXFEL compatible with high pressure liquid delivery of an SFX experiment. While achieving ~60% reduction in sample waste, we determine the structure of the enzyme 3-deoxy-D-manno-octulosonate-8-phosphate synthase from microcrystals delivered in droplets revealing distinct structural features not previously reported.
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Sep 2020
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I03-Macromolecular Crystallography
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Max O.
Wiedorn
,
Dominik
Oberthuer
,
Richard
Bean
,
Robin
Schubert
,
Nadine
Werner
,
Brian
Abbey
,
Martin
Aepfelbacher
,
Luigi
Adriano
,
Aschkan
Allahgholi
,
Nasser
Al-Qudami
,
Jakob
Andreasson
,
Steve
Aplin
,
Salah
Awel
,
Kartik
Ayyer
,
Saša
Bajt
,
Imrich
Barák
,
Sadia
Bari
,
Johan
Bielecki
,
Sabine
Botha
,
Djelloul
Boukhelef
,
Wolfgang
Brehm
,
Sandor
Brockhauser
,
Igor
Cheviakov
,
Matthew A.
Coleman
,
Francisco
Cruz-Mazo
,
Cyril
Danilevski
,
Connie
Darmanin
,
R. Bruce
Doak
,
Martin
Domaracky
,
Katerina
Dörner
,
Yang
Du
,
Hans
Fangohr
,
Holger
Fleckenstein
,
Matthias
Frank
,
Petra
Fromme
,
Alfonso M.
Gañán-Calvo
,
Yaroslav
Gevorkov
,
Klaus
Giewekemeyer
,
Helen Mary
Ginn
,
Heinz
Graafsma
,
Rita
Graceffa
,
Dominic
Greiffenberg
,
Lars
Gumprecht
,
Peter
Göttlicher
,
Janos
Hajdu
,
Steffen
Hauf
,
Michael
Heymann
,
Susannah
Holmes
,
Daniel A.
Horke
,
Mark S.
Hunter
,
Siegfried
Imlau
,
Alexander
Kaukher
,
Yoonhee
Kim
,
Alexander
Klyuev
,
Juraj
Knoška
,
Bostjan
Kobe
,
Manuela
Kuhn
,
Christopher
Kupitz
,
Jochen
Küpper
,
Janine Mia
Lahey-Rudolph
,
Torsten
Laurus
,
Karoline
Le Cong
,
Romain
Letrun
,
P. Lourdu
Xavier
,
Luis
Maia
,
Filipe R. N. C.
Maia
,
Valerio
Mariani
,
Marc
Messerschmidt
,
Markus
Metz
,
Davide
Mezza
,
Thomas
Michelat
,
Grant
Mills
,
Diana C. F.
Monteiro
,
Andrew
Morgan
,
Kerstin
Mühlig
,
Anna
Munke
,
Astrid
Münnich
,
Julia
Nette
,
Keith A.
Nugent
,
Theresa
Nuguid
,
Allen M.
Orville
,
Suraj
Pandey
,
Gisel
Pena
,
Pablo
Villanueva-Perez
,
Jennifer
Poehlsen
,
Gianpietro
Previtali
,
Lars
Redecke
,
Winnie Maria
Riekehr
,
Holger
Rohde
,
Adam
Round
,
Tatiana
Safenreiter
,
Iosifina
Sarrou
,
Tokushi
Sato
,
Marius
Schmidt
,
Bernd
Schmitt
,
Robert
Schönherr
,
Joachim
Schulz
,
Jonas A.
Sellberg
,
M. Marvin
Seibert
,
Carolin
Seuring
,
Megan L.
Shelby
,
Robert L.
Shoeman
,
Marcin
Sikorski
,
Alessandro
Silenzi
,
Claudiu A.
Stan
,
Xintian
Shi
,
Stephan
Stern
,
Jola
Sztuk-Dambietz
,
Janusz
Szuba
,
Aleksandra
Tolstikova
,
Martin
Trebbin
,
Ulrich
Trunk
,
Patrik
Vagovic
,
Thomas
Ve
,
Britta
Weinhausen
,
Thomas A.
White
,
Krzysztof
Wrona
,
Chen
Xu
,
Oleksandr
Yefanov
,
Nadia
Zatsepin
,
Jiaguo
Zhang
,
Markus
Perbandt
,
Adrian P.
Mancuso
,
Christian
Betzel
,
Henry
Chapman
,
Anton
Barty
Open Access
Abstract: The new European X-ray Free-Electron Laser is the first X-ray free-electron laser capable of delivering X-ray pulses with a megahertz inter-pulse spacing, more than four orders of magnitude higher than previously possible. However, to date, it has been unclear whether it would indeed be possible to measure high-quality diffraction data at megahertz pulse repetition rates. Here, we show that high-quality structures can indeed be obtained using currently available operating conditions at the European XFEL. We present two complete data sets, one from the well-known model system lysozyme and the other from a so far unknown complex of a β-lactamase from K. pneumoniae involved in antibiotic resistance. This result opens up megahertz serial femtosecond crystallography (SFX) as a tool for reliable structure determination, substrate screening and the efficient measurement of the evolution and dynamics of molecular structures using megahertz repetition rate pulses available at this new class of X-ray laser source.
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Oct 2018
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Abstract: A quantitative height profile image of a silicon nano-trench structure was obtained via coherent diffraction imaging (CDI) utilizing focused X-rays at a photon energy of 5.5 keV. The ability to optimize the spatial coherence and the photon flux density of a focused X-ray beam was the key technique for achieving such technical progress at a given X-ray photon flux. This was achieved by investigating the tunability of the focused beam’s optical properties and performing a CDI experiment with the focused X-rays. The relationship between the focused X-rays’ optical properties (e.g., photon flux density and spatial coherence length) and the incident beam’s size, which can be tuned by adjusting the slits in front of the Fresnel zone plate (FZP) was elucidated. We also obtained a quantitative image of a nano-trench sample produced via the reconstruction process of CDI, which utilizes carefully tuned, focused X-rays.
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May 2016
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