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Article ; Online: Multiscale Biofabrication: Integrating Additive Manufacturing with DNA-Programmable Self-Assembly.

Jäkel, Anna C / Heymann, Michael / Simmel, Friedrich C

Advanced biology

2022 Volume 7, Issue 3, Page(s) e2200195

Abstract: Structure and hierarchical organization are crucial elements of biological systems and are likely required when engineering synthetic biomaterials with life-like behavior. In this context, additive manufacturing techniques like bioprinting have become ... ...

Abstract	Structure and hierarchical organization are crucial elements of biological systems and are likely required when engineering synthetic biomaterials with life-like behavior. In this context, additive manufacturing techniques like bioprinting have become increasingly popular. However, 3D bioprinting, as well as other additive manufacturing techniques, show limited resolution, making it difficult to yield structures on the sub-cellular level. To be able to form macroscopic synthetic biological objects with structuring on this level, manufacturing techniques have to be used in conjunction with biomolecular nanotechnology. Here, a short overview of both topics and a survey of recent advances to combine additive manufacturing with microfabrication techniques and bottom-up self-assembly involving DNA, are given.
MeSH term(s)	Biocompatible Materials ; Microtechnology ; Bioprinting/methods ; Nanotechnology
Chemical Substances	Biocompatible Materials
Language	English
Publishing date	2022-11-03
Publishing country	Germany
Document type	Journal Article ; Review ; Research Support, Non-U.S. Gov't
ISSN	2701-0198
ISSN (online)	2701-0198
DOI	10.1002/adbi.202200195
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Article ; Online: Helium-electrospray improves sample delivery in X-ray single-particle imaging experiments.

Yenupuri, Tej Varma / Rafie-Zinedine, Safi / Worbs, Lena / Heymann, Michael / Schulz, Joachim / Bielecki, Johan / Maia, Filipe R N C

Scientific reports

2024 Volume 14, Issue 1, Page(s) 4401

Abstract: Imaging the structure and observing the dynamics of isolated proteins using single-particle X-ray diffractive imaging (SPI) is one of the potential applications of X-ray free-electron lasers (XFELs). Currently, SPI experiments on isolated proteins are ... ...

Abstract	Imaging the structure and observing the dynamics of isolated proteins using single-particle X-ray diffractive imaging (SPI) is one of the potential applications of X-ray free-electron lasers (XFELs). Currently, SPI experiments on isolated proteins are limited by three factors: low signal strength, limited data and high background from gas scattering. The last two factors are largely due to the shortcomings of the aerosol sample delivery methods in use. Here we present our modified electrospray ionization (ESI) source, which we dubbed helium-ESI (He-ESI). With it, we increased particle delivery into the interaction region by a factor of 10, for 26 nm-sized biological particles, and decreased the gas load in the interaction chamber corresponding to an 80% reduction in gas scattering when compared to the original ESI. These improvements have the potential to significantly increase the quality and quantity of SPI diffraction patterns in future experiments using He-ESI, resulting in higher-resolution structures.
MeSH term(s)	Helium ; X-Rays ; X-Ray Diffraction ; Proteins ; Lasers
Chemical Substances	Helium (206GF3GB41) ; Proteins
Language	English
Publishing date	2024-02-22
Publishing country	England
Document type	Journal Article
ZDB-ID	2615211-3
ISSN	2045-2322 ; 2045-2322
ISSN (online)	2045-2322
ISSN	2045-2322
DOI	10.1038/s41598-024-54605-9
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Article ; Online: Enhancing electrospray ionization efficiency for particle transmission through an aerodynamic lens stack.

Rafie-Zinedine, Safi / Varma Yenupuri, Tej / Worbs, Lena / Maia, Filipe R N C / Heymann, Michael / Schulz, Joachim / Bielecki, Johan

Journal of synchrotron radiation

2024 Volume 31, Issue Pt 2, Page(s) 222–232

Abstract: This work investigates the performance of the electrospray aerosol generator at the European X-ray Free Electron Laser (EuXFEL). This generator is, together with an aerodynamic lens stack that transports the particles into the X-ray interaction vacuum ... ...

Abstract	This work investigates the performance of the electrospray aerosol generator at the European X-ray Free Electron Laser (EuXFEL). This generator is, together with an aerodynamic lens stack that transports the particles into the X-ray interaction vacuum chamber, the method of choice to deliver particles for single-particle coherent diffractive imaging (SPI) experiments at the EuXFEL. For these experiments to be successful, it is necessary to achieve high transmission of particles from solution into the vacuum interaction region. Particle transmission is highly dependent on efficient neutralization of the charged aerosol generated by the electrospray mechanism as well as the geometry in the vicinity of the Taylor cone. We report absolute particle transmission values for different neutralizers and geometries while keeping the conditions suitable for SPI experiments. Our findings reveal that a vacuum ultraviolet ionizer demonstrates a transmission efficiency approximately seven times greater than the soft X-ray ionizer used previously. Combined with an optimized orifice size on the counter electrode, we achieve >40% particle transmission from solution into the X-ray interaction region. These findings offer valuable insights for optimizing electrospray aerosol generator configurations and data rates for SPI experiments.
Language	English
Publishing date	2024-02-02
Publishing country	United States
Document type	Journal Article
ZDB-ID	2021413-3
ISSN	1600-5775 ; 0909-0495
ISSN (online)	1600-5775
ISSN	0909-0495
DOI	10.1107/S1600577524000158
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Article ; Online: Microfluidic characterization of macromolecular liquid-liquid phase separation.

Bremer, Anne / Mittag, Tanja / Heymann, Michael

Lab on a chip

2020 Volume 20, Issue 22, Page(s) 4225–4234

Abstract: Liquid-liquid phase separation plays important roles in the compartmentalization of cells. Developing an understanding of how phase separation is encoded in biomacromolecules requires quantitative mapping of their phase behavior. Given that such ... ...

Abstract	Liquid-liquid phase separation plays important roles in the compartmentalization of cells. Developing an understanding of how phase separation is encoded in biomacromolecules requires quantitative mapping of their phase behavior. Given that such experiments require large quantities of the biomolecule of interest, these efforts have been lagging behind the recent breadth of biological insights. Herein, we present a microfluidic phase chip that enables the measurement of saturation concentrations over at least three orders of magnitude for a broad spectrum of biomolecules and solution conditions. The phase chip consists of five units, each made of twenty individual sample chambers to allow the measurement of five sample conditions simultaneously. The analytes are slowly concentrated via evaporation of water, which is replaced by oil, until the sample undergoes phase separation into a dilute and dense phase. We show that the phase chip lowers the required sample quantity by 98% while offering six-fold better statistics in comparison to standard manual experiments that involve centrifugal separation of dilute and dense phases. We further show that the saturation concentrations measured in chips are in agreement with previously reported data for a variety of biomolecules. Concomitantly, time-dependent changes of the dense phase morphology and potential off-pathway processes, including aggregation, can be monitored microscopically. In summary, the phase chip is suited to exploring sequence-to-binodal relationships by enabling the determination of a large number of saturation concentrations at low protein cost.
MeSH term(s)	Microfluidics ; Water
Chemical Substances	Water (059QF0KO0R)
Language	English
Publishing date	2020-10-15
Publishing country	England
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	2056646-3
ISSN	1473-0189 ; 1473-0197
ISSN (online)	1473-0189
ISSN	1473-0197
DOI	10.1039/d0lc00613k
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Article ; Online: Membrane-coated 3D architectures for bottom-up synthetic biology.

Eto, Hiromune / Franquelim, Henri G / Heymann, Michael / Schwille, Petra

Soft matter

2021 Volume 17, Issue 22, Page(s) 5456–5466

Abstract: One of the great challenges of bottom-up synthetic biology is to recreate the cellular geometry and surface functionality required for biological reactions. Of particular interest are lipid membrane interfaces where many protein functions take place. ... ...

Abstract	One of the great challenges of bottom-up synthetic biology is to recreate the cellular geometry and surface functionality required for biological reactions. Of particular interest are lipid membrane interfaces where many protein functions take place. However, cellular 3D geometries are often complex, and custom-shaping stable lipid membranes on relevant spatial scales in the micrometer range has been hard to accomplish reproducibly. Here, we use two-photon direct laser writing to 3D print microenvironments with length scales relevant to cellular processes and reactions. We formed lipid bilayers on the surfaces of these printed structures, and we evaluated multiple combinatorial scenarios, where physiologically relevant membrane compositions were generated on several different polymer surfaces. Functional dynamic protein systems were reconstituted in vitro and their self-organization was observed in response to the 3D geometry. This method proves very useful to template biological membranes with an additional spatial dimension, and thus allows a better understanding of protein function in relation to the complex morphology of cells and organelles.
MeSH term(s)	Cell Membrane ; Lipid Bilayers ; Membranes ; Polymers ; Synthetic Biology
Chemical Substances	Lipid Bilayers ; Polymers
Language	English
Publishing date	2021-06-09
Publishing country	England
Document type	Journal Article
ZDB-ID	2191476-X
ISSN	1744-6848 ; 1744-683X
ISSN (online)	1744-6848
ISSN	1744-683X
DOI	10.1039/d1sm00112d
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Article ; Online: Fresnel lens optical fiber tweezers to evaluate the vitality of single algae cells.

Asadollahbaik, Asa / Kumar, Aashutosh / Heymann, Michael / Giessen, Harald / Fick, Jochen

Optics letters

2021 Volume 47, Issue 1, Page(s) 170–173

Abstract: Dunaliella salina algae are trapped and studied using dual-fiber optical tweezers based on nano-imprinted Fresnel lenses. Different forms of cyclic motion of living algae inside the optical trap are observed and analyzed. A characteristic periodic motion ...

Abstract	Dunaliella salina algae are trapped and studied using dual-fiber optical tweezers based on nano-imprinted Fresnel lenses. Different forms of cyclic motion of living algae inside the optical trap are observed and analyzed. A characteristic periodic motion in the 0-35 Hz frequency region reflects the algal flagella activity and is used to estimate the algal vitality, by photomovement. The trap stiffness and optical forces are measured for the case of a dead algal cell. It is shown that the dual-fiber optical tweezers can be used to study the vitality (or viability) property of single cells, a property that is essential and can be scaled up to other applications, such as sperm analysis for fertility tests.
MeSH term(s)	Motion ; Optical Fibers ; Optical Tweezers
Language	English
Publishing date	2021-12-24
Publishing country	United States
Document type	Journal Article
ISSN	1539-4794
ISSN (online)	1539-4794
DOI	10.1364/OL.447683
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Article ; Online: 3D printed protein-based robotic structures actuated by molecular motor assemblies.

Jia, Haiyang / Flommersfeld, Johannes / Heymann, Michael / Vogel, Sven K / Franquelim, Henri G / Brückner, David B / Eto, Hiromune / Broedersz, Chase P / Schwille, Petra

Nature materials

2022 Volume 21, Issue 6, Page(s) 703–709

Abstract: Upscaling motor protein activity to perform work in man-made devices has long been an ambitious goal in bionanotechnology. The use of hierarchical motor assemblies, as realized in sarcomeres, has so far been complicated by the challenges of arranging ... ...

Abstract	Upscaling motor protein activity to perform work in man-made devices has long been an ambitious goal in bionanotechnology. The use of hierarchical motor assemblies, as realized in sarcomeres, has so far been complicated by the challenges of arranging sufficiently high numbers of motor proteins with nanoscopic precision. Here, we describe an alternative approach based on actomyosin cortex-like force production, allowing low complexity motor arrangements in a contractile meshwork that can be coated onto soft objects and locally activated by ATP. The design is reminiscent of a motorized exoskeleton actuating protein-based robotic structures from the outside. It readily supports the connection and assembly of micro-three-dimensional printed modules into larger structures, thereby scaling up mechanical work. We provide an analytical model of force production in these systems and demonstrate the design flexibility by three-dimensional printed units performing complex mechanical tasks, such as microhands and microarms that can grasp and wave following light activation.
MeSH term(s)	Humans ; Printing, Three-Dimensional ; Robotic Surgical Procedures ; Robotics
Language	English
Publishing date	2022-05-26
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2088679-2
ISSN	1476-4660 ; 1476-1122
ISSN (online)	1476-4660
ISSN	1476-1122
DOI	10.1038/s41563-022-01258-6
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Article ; Online: Cell-free expression of RNA encoded genes using MS2 replicase.

Weise, Laura I / Heymann, Michael / Mayr, Viktoria / Mutschler, Hannes

Nucleic acids research

2019 Volume 47, Issue 20, Page(s) 10956–10967

Abstract: RNA replicases catalyse transcription and replication of viral RNA genomes. Of particular interest for in vitro studies are phage replicases due to their small number of host factors required for activity and their ability to initiate replication in the ... ...

Abstract	RNA replicases catalyse transcription and replication of viral RNA genomes. Of particular interest for in vitro studies are phage replicases due to their small number of host factors required for activity and their ability to initiate replication in the absence of any primers. However, the requirements for template recognition by most phage replicases are still only poorly understood. Here, we show that the active replicase of the archetypical RNA phage MS2 can be produced in a recombinant cell-free expression system. We find that the 3' terminal fusion of antisense RNAs with a domain derived from the reverse complement of the wild type MS2 genome generates efficient templates for transcription by the MS2 replicase. The new system enables DNA-independent gene expression both in batch reactions and in microcompartments. Finally, we demonstrate that MS2-based RNA-dependent transcription-translation reactions can be used to control DNA-dependent gene expression by encoding a viral DNA-dependent RNA polymerase on a MS2 RNA template. Our study sheds light on the template requirements of the MS2 replicase and paves the way for new in vitro applications including the design of genetic circuits combining both DNA- and RNA-encoded systems.
MeSH term(s)	Cell-Free System ; Emulsions/chemistry ; Genes, Viral ; Levivirus/enzymology ; Levivirus/genetics ; Protein Biosynthesis ; Protein Subunits/genetics ; RNA, Viral/genetics ; RNA-Dependent RNA Polymerase/genetics ; Transcription, Genetic
Chemical Substances	Emulsions ; Protein Subunits ; RNA, Viral ; RNA-Dependent RNA Polymerase (EC 2.7.7.48)
Language	English
Publishing date	2019-09-30
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	186809-3
ISSN	1362-4962 ; 1362-4954 ; 0301-5610 ; 0305-1048
ISSN (online)	1362-4962 ; 1362-4954
ISSN	0301-5610 ; 0305-1048
DOI	10.1093/nar/gkz817
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Article ; Online: Temperature-sensitive protein expression in protocells.

Jia, Haiyang / Heymann, Michael / Härtel, Tobias / Kai, Lei / Schwille, Petra

Chemical communications (Cambridge, England)

2019 Volume 55, Issue 45, Page(s) 6421–6424

Abstract: We engineered a synthetic temperature regulation toolbox to enable protocells to sense and respond to heat, utilizing RNA thermometers. The thermo-sensitive protocells were generated by encapsulating temperature feedback transcription/translation ... ...

Abstract	We engineered a synthetic temperature regulation toolbox to enable protocells to sense and respond to heat, utilizing RNA thermometers. The thermo-sensitive protocells were generated by encapsulating temperature feedback transcription/translation machinery in droplets. Based on these temperature-sensing devices, the protocells can be operated with logic AND gates, differentially processing temperature stimuli into biological signals.
MeSH term(s)	Emulsions/chemistry ; Particle Size ; Proteins/genetics ; Proteins/metabolism ; RNA/metabolism ; Temperature
Chemical Substances	Emulsions ; Proteins ; RNA (63231-63-0)
Language	English
Publishing date	2019-05-16
Publishing country	England
Document type	Journal Article
ZDB-ID	1472881-3
ISSN	1364-548X ; 1359-7345 ; 0009-241X
ISSN (online)	1364-548X
ISSN	1359-7345 ; 0009-241X
DOI	10.1039/c9cc02734c
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Book: Nanoskaliger Kieselsaeureschlamm - Technologieentwicklungen zur Verwertung in mineralischen Baustoffen mit dem Ziel der Verbesserung der Werkstoffeigenschaften

Heymann, Michael

Title variant	Nanoscale silicic acid sludge: Technology developments for utilization in mineral building materials with the aim of enhancing the properties of materials
Institution	Brueckner Grundbau GmbH, Niederlassung Sued-Ost, Cottaer Str. 2, 01159, Dresden, DE
Keywords	Abwasserbehandlung ; Industrieabwasser ; Reststoff ; Baustoff ; Werkstoffkunde ; Baustelle ; Bauwerk ; Nanomaterialien ; Schlammverwertung ; Stoffliche Verwertung ; Sekundaerrohstoff ; Abfallverwertung ; Anorganische Siliziumverbindung ; Tiefbau ; Eignungspruefung ; Schlamm ; Recyclingprodukt ; Bauwirtschaft
Document type	Book
Remark	project start: 05/01/2010 project end: 04/30/2012 grant ID: 03X0081K
Database	Environmental research database (UFORDAT) of the German Federal Environment Agency (UBA)

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