* joint first author # joint corresponding author

Weida Liao, Elena Erben, Moritz Kreysing, Eric Lauga
Theoretical model of confined thermoviscous flows for artificial cytoplasmic streaming.
Phys Rev Fluids, 8 Art. No. 034202 (2023)
Recent experiments in cell biology have probed the impact of artificially induced intracellular flows in the spatiotemporal organization of cells and organisms. In these experiments, mild dynamical heating (a few kelvins) via focused infrared light from a laser leads to long-range, thermoviscous flows of the cytoplasm inside a cell. To extend future use of this method in cell biology, popularized as focused-light-induced cytoplasmic streaming (FLUCS), new quantitative models are needed to link the external light forcing to the produced flows and transport. Here, we present a fully analytical, theoretical model describing the fluid flow induced by the dynamical laser stimulus at all length scales (both near the scan path of the laser beam and in the far field) in two-dimensional confinement. We model the effect of the focused light as a small, local temperature change in the fluid, which causes a small change in both the density and the viscosity of the fluid locally. In turn, this results in a locally compressible fluid flow. We analytically solve for the instantaneous flow field induced by the translation of a heat spot of arbitrary timedependent amplitude along a scan path of arbitrary length. We show that the leading-order instantaneous flow field results from the thermal expansion of the fluid and is independent of the thermal viscosity coefficient. This leading-order velocity field is proportional to the thermal expansion coefficient and the magnitude of the temperature perturbation, with far-field behavior typically dominated by a source or sink flow and proportional to the rate of change of the heat-spot amplitude. In contrast, and in agreement with experimental measurements, the net displacement of a material point due to a full scan of the heat spot is quadratic in the heat-spot amplitude, as it results from the interplay of thermal expansion and thermal viscosity changes. The corresponding average velocity of material points over a scan is a hydrodynamic source dipole in the far field, with direction dependent on the relative importance of thermal expansion and thermal viscosity changes. Our quantitative findings show excellent agreement with recent experimental results and will enable the design of new controlled experiments to establish the physiological role of physical transport processes inside cells.

Iliya D. Stoev, Florian Huhn, Nicola Maghelli, Elena Erben, Sebastian T. Bundschuh, Benjamin Seelbinder, Catarina Nabais, Britta Schroth-Diez, Ines Höfer, Jan Peychl, Moritz Kreysing, Gert Rapp
FLUCS - Focused Light-Induced Cytoplasmic Streaming : Optically controlled microflows for biology and the life sciences.
Imaging & Microscopy, 24(2) 22-24 (2022)
Open Access
FLUCS is a new photomanipulation technique that allows scientists to optically generate and precisely control microscopic flows within cells, embryos and other viscous fluids. The development of the module is the result of a collaboration between engineers and scientists from Rapp OptoElectronic, the Max Planck Institute of Molecular Cell Biology and Genetics (MPI-CBG) in Dresden, Germany, and Olympus Europe. The integration of the Rapp FLUCS module into the Olympus IXplore SpinSR spinning disk confocal microscope system allows to generate microflows in the specimen and to image the resulting dynamic biological processes at high spatial and temporal resolution. The system is now available as a finalized and tested product to a broad scientific community worldwide. https://analyticalscience.wiley.com/do/10.1002/was.0004000255; https://analyticalscience.wiley.com/do/10.1002/was.00070072/full/blaetterkatalog_sim0222.pdf

Adriano Acunzo, Emanuela Scardapane, Maria De Luca, Daniele Marra, Raffaele Velotta, Antonio Minopoli
Plasmonic Nanomaterials for Colorimetric Biosensing: A Review.
Chemosensors, 10(4) Art. No. 136 (2022)
Open Access DOI
In the last few decades, plasmonic colorimetric biosensors raised increasing interest in bioanalytics thanks to their cost-effectiveness, responsiveness, and simplicity as compared to conventional laboratory techniques. Potential high-throughput screening and easy-to-use assay procedures make them also suitable for realizing point of care devices. Nevertheless, several challenges such as fabrication complexity, laborious biofunctionalization, and poor sensitivity compromise their technological transfer from research laboratories to industry and, hence, still hamper their adoption on large-scale. However, newly-developing plasmonic colorimetric biosensors boast impressive sensing performance in terms of sensitivity, dynamic range, limit of detection, reliability, and specificity thereby continuously encouraging further researches. In this review, recently reported plasmonic colorimetric biosensors are discussed with a focus on the following categories: (i) on-platform-based (localized surface plasmon resonance, coupled plasmon resonance and surface lattice resonance); (ii) colloid aggregation-based (label-based and label free); (iii) colloid non-aggregation-based (nanozyme, etching-based and growth-based).

Juan M Iglesias-Artola, Björn Drobot, Mrityunjoy Kar, Anatol Fritsch, Hannes Mutschler, T-Y Dora Tang, Moritz Kreysing
Charge-density reduction promotes ribozyme activity in RNA-peptide coacervates via RNA fluidization and magnesium partitioning.
Nat Chem, 14(4) 407-416 (2022)
Open Access PDF DOI
It has long been proposed that phase-separated compartments can provide a basis for the formation of cellular precursors in prebiotic environments. However, we know very little about the properties of coacervates formed from simple peptides, their compatibility with ribozymes or their functional significance. Here we assess the conditions under which functional ribozymes form coacervates with simple peptides. We find coacervation to be most robust when transitioning from long homopeptides to shorter, more pre-biologically plausible heteropeptides. We mechanistically show that these RNA-peptide coacervates display peptide-dependent material properties and cofactor concentrations. We find that the interspacing of cationic and neutral amino acids increases RNA mobility, and we use isothermal calorimetry to reveal sequence-dependent Mg2+ partitioning, two critical factors that together enable ribozyme activity. Our results establish how peptides of limited length, homogeneity and charge density facilitate the compartmentalization of active ribozymes into non-gelating, magnesium-rich coacervates, a scenario that could be applicable to cellular precursors with peptide-dependent functional phenotypes.

Benjamin Seelbinder, Manavi Jain, Elena Erben, Sergei Klykov, Iliya D. Stoev, Moritz Kreysing
Non-invasive Chromatin Deformation and Measurement of Differential Mechanical Properties in the Nucleus.
bioRxiv, Art. No. https://doi.org/10.1101/2021.12.15.472786 (2021)
Open Access PDF DOI
The nucleus is highly organized to facilitate coordinated gene transcription. Measuring the rheological properties of the nucleus and its sub-compartments will be crucial to understand the principles underlying nuclear organization. Here, we show that strongly localized temperature gradients (approaching 1°C /μm) can lead to substantial intra-nuclear chromatin displacements (>1 μm), while nuclear area and lamina shape remain unaffected. Using particle image velocimetry (PIV), intra-nuclear displacement fields can be calculated and converted into spatio-temporally resolved maps of various strain components. Using this approach, we show that chromatin displacements are highly reversible, indicating that elastic contributions are dominant in maintaining nuclear organization on the time scale of seconds. In genetically inverted nuclei, centrally compacted heterochromatin displays high resistance to deformation, giving a rigid, solid-like appearance. Correlating spatially resolved strain maps with fluorescent reporters in conventional interphase nuclei reveals that various nuclear compartments possess distinct mechanical identities. Surprisingly, both densely and loosely packed chromatin showed high resistance to deformation, compared to medium dense chromatin. Equally, nucleoli display particularly high rigidity and strong local anchoring to heterochromatin. Our results establish how localized temperature gradients can be used to drive nuclear compartments out of mechanical equilibrium to obtain spatial maps of their material responses.

Iliya D. Stoev, Benjamin Seelbinder, Elena Erben, Nicola Maghelli, Moritz Kreysing
Highly sensitive force measurements in an optically generated, harmonic hydrodynamic trap.
eLight, 1 Art. No. 7 (2021)
Open Access PDF DOI
The use of optical tweezers to measure forces acting upon microscopic particles has revolutionised fields from material science to cell biology. However, despite optical control capabilities, this technology is highly constrained by the material properties of the probe, and its use may be limited due to concerns about the effect on biological processes. Here we present a novel, optically controlled trapping method based on light-induced hydrodynamic flows. Specifically, we leverage optical control capabilities to convert a translationally invariant topological defect of a flow field into an attractor for colloids in an effectively one-dimensional harmonic, yet freely rotatable system. Circumventing the need to stabilise particle dynamics along an unstable axis, this novel trap closely resembles the isotropic dynamics of optical tweezers. Using magnetic beads, we explicitly show the existence of a linear force-extension relationship that can be used to detect femtoNewton-range forces with sensitivity close to the thermal limit. Our force measurements remove the need for laser-particle contact, while also lifting material constraints, which renders them a particularly interesting tool for the life sciences and engineering.

Sara N. Nagelberg✳︎, Jan F. Totz✳︎, Matthäus Mittasch, Vishnu Sresht, Lukas Zeininger, Timothy M Swager, Moritz Kreysing, Mathias Kolle
Actuation of Janus Emulsion Droplets via Optothermally Induced Marangoni Forces.
Phys Rev Lett, 127(14) Art. No. 144503 (2021)
Microscale Janus emulsions represent a versatile material platform for dynamic refractive, reflective, and light-emitting optical components. Here, we present a mechanism for droplet actuation that exploits thermocapillarity. Using optically induced thermal gradients, an interfacial tension differential is generated across the surfactant-free internal capillary interface of Janus droplets. The interfacial tension differential causes droplet-internal Marangoni flows and a net torque, resulting in a predictable and controllable reorientation of the droplets. The effect can be quantitatively described with a simple model that balances gravitational and thermal torques. Occurring in small thermal gradients, these optothermally induced Marangoni dynamics represent a promising mechanism for controlling droplet-based micro-optical components.

Anatol Fritsch✳︎, Andrés F Diaz-Delgadillo✳︎, Omar Adame-Arana✳︎, Carsten Hoege, Matthäus Mittasch, Moritz Kreysing, Mark Leaver, Anthony Hyman, Frank Jülicher, Christoph A. Weber
Local thermodynamics govern formation and dissolution of Caenorhabditis elegans P granule condensates.
Proc Natl Acad Sci U.S.A., 118(37) Art. No. e2102772118 (2021)
Membraneless compartments, also known as condensates, provide chemically distinct environments and thus spatially organize the cell. A well-studied example of condensates is P granules in the roundworm Caenorhabditis elegans that play an important role in the development of the germline. P granules are RNA-rich protein condensates that share the key properties of liquid droplets such as a spherical shape, the ability to fuse, and fast diffusion of their molecular components. An outstanding question is to what extent phase separation at thermodynamic equilibrium is appropriate to describe the formation of condensates in an active cellular environment. To address this question, we investigate the response of P granule condensates in living cells to temperature changes. We observe that P granules dissolve upon increasing the temperature and recondense upon lowering the temperature in a reversible manner. Strikingly, this temperature response can be captured by in vivo phase diagrams that are well described by a Flory-Huggins model at thermodynamic equilibrium. This finding is surprising due to active processes in a living cell. To address the impact of such active processes on intracellular phase separation, we discuss temperature heterogeneities. We show that, for typical estimates of the density of active processes, temperature represents a well-defined variable and that mesoscopic volume elements are at local thermodynamic equilibrium. Our findings provide strong evidence that P granule assembly and disassembly are governed by phase separation based on local thermal equilibria where the nonequilibrium nature of the cytoplasm is manifested on larger scales.

Elena Erben, Benjamin Seelbinder, Iliya D. Stoev, Sergei Klykov, Nicola Maghelli, Moritz Kreysing
Feedback-based positioning and diffusion suppression of particles via optical control of thermoviscous flows.
Opt express, 29(19) 30272-30283 (2021)
Open Access PDF DOI
The ability to control the position of micron-size particles with high precision using tools such as optical tweezers has led to major advances in fields such as biology, physics and material science. In this paper, we present a novel optical strategy to confine particles in solution with high spatial control using feedback-controlled thermoviscous flows. We show that this technique allows micron-size particles to be positioned and confined with subdiffraction precision (24 nm), effectively suppressing their diffusion. Due to its physical characteristics, our approach might be particular attractive where laser exposure is of concern or materials are inherently incompatible with optical tweezing since it does not rely on contrast in the refractive index.

Anatol W Fritsch✳︎, Andrés Diaz Delgadillo✳︎, Omar Adame-Arana✳︎, Carsten Hoege, Matthäus Mittasch, Moritz Kreysing, Mark Leaver, Anthony A. Hyman, Frank Jülicher, Christoph A. Weber
Local thermodynamics governs the formation and dissolution of protein condensates in living cells.
bioRxiv, Art. No. https://doi.org/10.1101/2021.02.11.430794 (2021)
Open Access PDF DOI

Nicolas T Chartier✳︎, Arghyadip Mukherjee✳︎, Julia Pfanzelter✳︎, Sebastian Fürthauer, Ben T Larson, Anatol Fritsch, Rana Amini, Moritz Kreysing, Frank Jülicher#, Stephan W. Grill#
A hydraulic instability drives the cell death decision in the nematode germline.
Nat Phys, 17(8) 920-925 (2021)
Open Access PDF DOI
Oocytes are large cells that develop into an embryo upon fertilization1. As interconnected germ cells mature into oocytes, some of them grow-typically at the expense of others that undergo cell death2-4. We present evidence that in the nematode Caenorhabditis elegans, this cell-fate decision is mechanical and related to tissue hydraulics. An analysis of germ cell volumes and material fluxes identifies a hydraulic instability that amplifies volume differences and causes some germ cells to grow and others to shrink, a phenomenon that is related to the two-balloon instability5. Shrinking germ cells are extruded and they die, as we demonstrate by artificially reducing germ cell volumes via thermoviscous pumping6. Our work reveals a hydraulic symmetry-breaking transition central to the decision between life and death in the nematode germline.

Kaushikaram Subramanian✳︎, Heike Petzold✳︎, Benjamin Seelbinder✳︎, Lena Hersemann, Ina Nüsslein, Moritz Kreysing
Optical plasticity of mammalian cells.
J Biophotonics, 14(4) Art. No. e202000457 (2020)
Open Access PDF DOI
Transparency is widespread in nature, ranging from transparent insect wings to ocular tissues that enable you to read this text, and transparent marine vertebrates. And yet, cells and tissue models in biology are usually strongly light scattering and optically opaque, precluding deep optical microscopy. Here we describe the directed evolution of cultured mammalian cells toward increased transparency. We find that mutations greatly diversify the optical phenotype of Chinese Hamster Ovary cells, a cultured mammalian cell line. Furthermore, only three rounds of high-throughput optical selection and competitive growth are required to yield fit cells with greatly improved transparency. Based on 15 monoclonal cell lines derived from this directed evolution experiment, we find that the evolved transparency frequently goes along with a reduction of nuclear granularity and physiological shifts in gene expression profiles. In the future this optical plasticity of mammalian cells may facilitate genetic clearance of living tissues for in vivo microscopy.

Juliana G. Roscito, Kaushikaram Subramanian, Ronald Naumann, Mihail Sarov, Anna Shevchenko, Aliona Bogdanova, Thomas Kurth, Leo Foerster, Moritz Kreysing, Michael Hiller
Recapitulating evolutionary divergence in a single cis-regulatory element is sufficient to cause expression changes of the lens gene Tdrd7.
Mol Biol Evol, 38(2) 380-392 (2020)
Open Access PDF DOI
Mutations in cis-regulatory elements play important roles for phenotypic changes during evolution. Eye degeneration in the blind mole rat (BMR; Nannospalax galili) and other subterranean mammals is significantly associated with widespread divergence of eye regulatory elements, but the effect of these regulatory mutations on eye development and function has not been explored. Here, we investigate the effect of mutations observed in the BMR sequence of a conserved non-coding element upstream of Tdrd7, a pleiotropic gene required for lens development and spermatogenesis. We first show that this conserved element is a transcriptional repressor in lens cells and that the BMR sequence partially lost repressor activity. Next, we recapitulated evolutionary changes in this element by precisely replacing the endogenous regulatory element in a mouse line by the orthologous BMR sequence with CRISPR-Cas9. Strikingly, this repressor replacement caused a more than two-fold up-regulation of Tdrd7 in the developing lens; however, increased mRNA level does not result in a corresponding increase in TDRD7 protein nor an obvious lens phenotype, possibly explained by buffering at the posttranscriptional level. Our results are consistent with eye degeneration in subterranean mammals having a polygenic basis where many small-effect mutations in different eye-regulatory elements collectively contribute to phenotypic differences.

Nicolas T Chartier, Arghyadip Mukherjee, Julia Pfannzelter, Sebastian Fürthauer, Ben T Larson, Anatol W Fritsch, Moritz Kreysing, Frank Jülicher, Stephan W. Grill
A hydraulic instability drives the cell death decision in the nematode germline
bioRxiv, Art. No. https://doi.org/10.1101/2020.05.30.125864 (2020)
Open Access PDF DOI
Oocytes are large and resourceful. During oogenesis some germ cells grow, typically at the expense of others that undergo apoptosis. How germ cells are selected to live or die out of a homogeneous population remains unclear. Here we show that this cell fate decision in C. elegans is mechanical and related to tissue hydraulics. Germ cells become inflated when the pressure inside them is lower than in the common cytoplasmic pool. This condition triggers a hydraulic instability which amplifies volume differences and causes some germ cells to grow and others to shrink. Shrinking germ cells are extruded and die, as we demonstrate by reducing germ cell volumes via thermoviscous pumping. Together, this reveals a robust mechanism of mechanochemical cell fate decision making in the germline.

Christiane Iserman, Christine Desroches Altamirano, Ceciel Jegers, Ulrike Friedrich, Taraneh Zarin, Anatol Fritsch, Matthäus Mittasch, Antonio Domingues, Lena Hersemann, Marcus Jahnel, Doris Richter, Ulf-Peter Guenther, Matthias W. Hentze, Alan M Moses, Anthony Hyman, Günter Kramer, Moritz Kreysing, Titus Franzmann, Simon Alberti
Condensation of Ded1p Promotes a Translational Switch from Housekeeping to Stress Protein Production.
Cell, 181(4) 818-831 (2020)
Open Access PDF DOI
Cells sense elevated temperatures and mount an adaptive heat shock response that involves changes in gene expression, but the underlying mechanisms, particularly on the level of translation, remain unknown. Here we report that, in budding yeast, the essential translation initiation factor Ded1p undergoes heat-induced phase separation into gel-like condensates. Using ribosome profiling and an in vitro translation assay, we reveal that condensate formation inactivates Ded1p and represses translation of housekeeping mRNAs while promoting translation of stress mRNAs. Testing a variant of Ded1p with altered phase behavior as well as Ded1p homologs from diverse species, we demonstrate that Ded1p condensation is adaptive and fine-tuned to the maximum growth temperature of the respective organism. We conclude that Ded1p condensation is an integral part of an extended heat shock response that selectively represses translation of housekeeping mRNAs to promote survival under conditions of severe heat stress.

Matthäus Mittasch, Vanna M. Tran, Manolo U. Rios, Anatol Fritsch, Stephen Enos, Beatriz Ferreira Gomes, Alec Bond, Moritz Kreysing#, Jeffrey Woodruff#
Regulated changes in material properties underlie centrosome disassembly during mitotic exit.
J Cell Biol, 219(4) Art. No. e201912036 (2020)
Open Access PDF DOI
Centrosomes must resist microtubule-mediated forces for mitotic chromosome segregation. During mitotic exit, however, centrosomes are deformed and fractured by those same forces, which is a key step in centrosome disassembly. How the functional material properties of centrosomes change throughout the cell cycle, and how they are molecularly tuned, remain unknown. Here, we used optically induced flow perturbations to determine the molecular basis of centrosome strength and ductility in C. elegans embryos. We found that both properties declined sharply at anaphase onset, long before natural disassembly. This mechanical transition required PP2A phosphatase and correlated with inactivation of PLK-1 (Polo kinase) and SPD-2 (Cep192). In vitro, PLK-1 and SPD-2 directly protected centrosome scaffolds from force-induced disassembly. Our results suggest that, before anaphase, PLK-1 and SPD-2 respectively confer strength and ductility to the centrosome scaffold so that it can resist microtubule-pulling forces. In anaphase, centrosomes lose PLK-1 and SPD-2 and transition to a weak, brittle state that enables force-mediated centrosome disassembly.

Kaushikaram Subramanian, Martin Weigert, Oliver Borsch, Heike Petzold, Alfonso Garcia-Ulloa, Eugene W Myers, Marius Ader, Irina Solovei, Moritz Kreysing
Rod nuclear architecture determines contrast transmission of the retina and behavioral sensitivity in mice.
Elife, 8 Art. No. e49542 (2019)
Open Access PDF DOI
Rod photoreceptors of nocturnal mammals display a striking inversion of nuclear architecture, which has been proposed as an evolutionary adaptation to dark environments. However, the nature of visual benefits and the underlying mechanisms remains unclear. It is widely assumed that improvements in nocturnal vision would depend on maximization of photon capture at the expense of image detail. Here we show that retinal optical quality improves 2-fold during terminal development, and that this enhancement is caused by nuclear inversion. We further demonstrate that improved retinal contrast transmission, rather than photon-budget or resolution, enhances scotopic contrast sensitivity by 18-27%, and improves motion detection capabilities up to 10-fold in dim environments. Our findings therefore add functional significance to a prominent exception of nuclear organization and establish retinal contrast transmission as a decisive determinant of mammalian visual perception.

Matthäus Mittasch, Vanna M. Tran, Manolo U. Rios, Anatol W Fritsch, Stephen Enos, Beatriz Ferreira Gomes, Alec Bond, Moritz Kreysing#, Jeffrey Woodruff#
Material aging causes centrosome weakening and disassembly during mitotic exit.
bioRxiv, Art. No. https://doi.org/10.1101/866434 (2019)
Open Access PDF DOI
Centrosomes must resist microtubule-mediated forces for mitotic chromosome segregation. During mitotic exit, however, centrosomes are deformed and fractured by those same forces, which is a key step in centrosome disassembly. How the functional material properties of centrosomes change throughout the cell cycle, and how they are molecularly tuned remain unknown. Here, we used optically-induced flow perturbations to determine the molecular basis of centrosome strength and ductility in C. elegans embryos. We found that both properties declined sharply at anaphase onset, long before natural disassembly. This mechanical transition required PP2A phosphatase and correlated with inactivation of PLK-1 (Polo Kinase) and SPD-2 (Cep192). In vitro, PLK-1 and SPD-2 directly protected centrosome scaffolds from force-induced disassembly. Our results suggest that, prior to anaphase, PLK-1 and SPD-2 confer strength and ductility to the centrosome scaffold so that it can resist microtubule-pulling forces. In anaphase, centrosomes lose PLK-1 and SPD-2 and transition to a weak, brittle state that enables force-mediated centrosome disassembly.

Kaushikaram Subramanian, Martin Weigert, Oliver Borsch, Heike Petzold, Alfonso Garcia, Eugene Myers, Marius Ader, Irina Solovei, Moritz Kreysing
Rod nuclear architecture determines contrast transmission of the retina and behavioral sensitivity in mice
bioRxiv, Art. No. https://doi.org/10.1101/752444 (2019)
Open Access PDF DOI
Rod photoreceptors of nocturnal mammals display a striking inversion of nuclear architecture, which has been proposed as an evolutionary adaptation to dark environments. However, the nature of visual benefits and underlying mechanisms remains unclear. It is widely assumed that improvements in nocturnal vision would depend on maximization of photon capture, at the expense of image detail. Here we show that retinal optical quality improves 2-fold during terminal development, which, confirmed by a mouse model, happens due to nuclear inversion. We further reveal that improved retinal contrast-transmission, rather than photon-budget or resolution, leads to enhanced contrast sensitivity under low light condition. Our findings therefore add functional significance to a prominent exception of nuclear organization and establish retinal contrast-transmission as a decisive determinant of mammalian visual perception

Moritz Kreysing
Probing the Functional Role of Physical Motion in Development.
Dev Cell, 51(2) 135-144 (2019)
Spatiotemporal organization during development has frequently been proposed to be explainable by reaction-transport models, where biochemical reactions couple to physical motion. However, whereas genetic tools allow causality of molecular players to be dissected via perturbation experiments, the functional role of physical transport processes, such as diffusion and cytoplasmic streaming, frequently remains untestable. This Perspective explores the challenges of validating reaction-transport hypotheses and highlights new opportunities provided by perturbation approaches that specifically target physical transport mechanisms. Using these methods, experimental physics may begin to catch up with molecular biology and find ways to test roles of diffusion and flows in development.

Kaushikaram Subramanian
Optics of the mammalian retina: Role of rod nuclear architecture in retinal transparency & visual sensitivity.
Ph.D. Thesis,Technische Universität Dresden, Dresden, Germany (2019)

Makoto Saito, Daniel Hess, Jan Eglinger, Anatol Fritsch, Moritz Kreysing, Brian Weinert, Chunaram Choudhary, Patrick Matthias
Acetylation of intrinsically disordered regions regulates phase separation.
Nat Chem Biol, 15(1) 51-61 (2019)
Liquid-liquid phase separation (LLPS) of proteins containing intrinsically disordered regions (IDRs) has been proposed as a mechanism underlying the formation of membrane-less organelles. Tight regulation of IDR behavior is essential to ensure that LLPS only takes place when necessary. Here, we report that IDR acetylation/deacetylation regulates LLPS and assembly of stress granules (SGs), membrane-less organelles forming in response to stress. Acetylome analysis revealed that the RNA helicase DDX3X, an important component of SGs, is a novel substrate of the deacetylase HDAC6. The N-terminal IDR of DDX3X (IDR1) can undergo LLPS in vitro, and its acetylation at multiple lysine residues impairs the formation of liquid droplets. We also demonstrated that enhanced LLPS propensity through deacetylation of DDX3X-IDR1 by HDAC6 is necessary for SG maturation, but not initiation. Our analysis provides a mechanistic framework to understand how acetylation and deacetylation of IDRs regulate LLPS spatiotemporally, and impact membrane-less organelle formation in vivo.

Björn Drobot, Juan M Iglesias-Artola, Kristian Le Vay, Viktoria Mayr, Mrityunjoy Kar, Moritz Kreysing#, Hannes Mutschler#, T-Y Dora Tang#
Compartmentalised RNA catalysis in membrane-free coacervate protocells.
Nat Commun, 9(1) Art. No. 3643 (2018)
Open Access PDF DOI
Phase separation of mixtures of oppositely charged polymers provides a simple and direct route to compartmentalisation via complex coacervation, which may have been important for driving primitive reactions as part of the RNA world hypothesis. However, to date, RNA catalysis has not been reconciled with coacervation. Here we demonstrate that RNA catalysis is viable within coacervate microdroplets and further show that these membrane-free droplets can selectively retain longer length RNAs while permitting transfer of lower molecular weight oligonucleotides.

Matthäus Mittasch
Light-driven intracellular flow perturbations reveal physical principles of cell organization.
Ph.D. Thesis,Technische Universität Dresden, Dresden, Germany (2018)

Matthäus Mittasch, Anatol Fritsch, Michael Nestler, Juan M Iglesias Artola, Kaushikaram Subramanian, Heike Petzold, Mrityunjoy Kar, Axel Voigt, Moritz Kreysing
Active gelation breaks time-reversal-symmetry of mitotic chromosome mechanics
bioRxiv, Art. No. https://doi.org/10.1101/296566 (2018)
Open Access PDF DOI
In cell division, mitosis is the phase in which duplicated sets of chromosomes are mechanically aligned to form the metaphase plate before being segregated in two daughter cells. Irreversibility is a hallmark of this process, despite the fundamental laws of Newtonian mechanics being time symmetric. Here we show experimentally that mitotic chromosomes receive the arrow of time by time-reversal-symmetry breaking of the underlying mechanics in prometaphase. By optically inducing hydrodynamic flows within prophase nuclei, we find that duplicated chromatid pairs initially form a fluid suspension in the nucleoplasm: although showing little motion on their own, condensed chromosomes are free to move through the nucleus in a time-reversible manner. Actively probing chromosome mobility further in time, we find that this viscous suspension of chromatin transitions into a gel after nuclear breakdown. This gel state, in which chromosomes cannot be moved by flows, persists even when chromosomes start moving to form the metaphase plate. Complemented by minimal reconstitution experiments, our active intra-nuclear micro-rheology reveals time-reversal-symmetry breaking of chromosome mechanics to be caused by the transition from a purely fluid suspension into an active gel.

Martin Weigert, Kaushikaram Subramanian, Sebastian T. Bundschuh, Eugene W Myers, Moritz Kreysing
Biobeam-Multiplexed wave-optical simulations of light-sheet microscopy.
PLoS Comput Biol, 14(4) Art. No. e1006079 (2018)
Open Access PDF DOI
Sample-induced image-degradation remains an intricate wave-optical problem in light-sheet microscopy. Here we present biobeam, an open-source software package that enables simulation of operational light-sheet microscopes by combining data from 105-106 multiplexed and GPU-accelerated point-spread-function calculations. The wave-optical nature of these simulations leads to the faithful reproduction of spatially varying aberrations, diffraction artifacts, geometric image distortions, adaptive optics, and emergent wave-optical phenomena, and renders image-formation in light-sheet microscopy computationally tractable.

Matthäus Mittasch, Peter Gross, Michael Nestler, Anatol W Fritsch, Christiane Iserman, Mrityunjoy Kar, Matthias Munder, Axel Voigt, Simon Alberti, Stephan W. Grill, Moritz Kreysing
Non-invasive perturbations of intracellular flow reveal physical principles of cell organization.
Nat Cell Biol, 20(3) 344-351 (2018)
Recent advances in cell biology enable precise molecular perturbations. The spatiotemporal organization of cells and organisms, however, also depends on physical processes such as diffusion or cytoplasmic flows, and strategies to perturb physical transport inside cells are not yet available. Here, we demonstrate focused-light-induced cytoplasmic streaming (FLUCS). FLUCS is local, directional, dynamic, probe-free, physiological, and is even applicable through rigid egg shells or cell walls. We explain FLUCS via time-dependent modelling of thermoviscous flows. Using FLUCS, we demonstrate that cytoplasmic flows drive partitioning-defective protein (PAR) polarization in Caenorhabditis elegans zygotes, and that cortical flows are sufficient to transport PAR domains and invert PAR polarity. In addition, we find that asymmetric cell division is a binary decision based on gradually varying PAR polarization states. Furthermore, the use of FLUCS for active microrheology revealed a metabolically induced fluid-to-solid transition of the yeast cytoplasm. Our findings establish how a wide range of transport-dependent models of cellular organization become testable by FLUCS.

Matthäus Mittasch, Peter Gross, Michael Nestler, Anatol W Fritsch, Christiane Iserman, Mrityunjoy Kar, Matthias Munder, Axel Voigt, Simon Alberti, Stephan W. Grill, Moritz Kreysing
How to apply FLUCS in single cells and living embryos.
Protocol Exchange, Art. No. doi:10.1038/protex.2017.157 (2018)
Open Access PDF DOI

Christof Mast, Friederike Möller, Moritz Kreysing, Severin Schink, Benedikt Obermayer, Ulrich Gerland, Dieter Braun
Toward Living Nanomachines
In: Living machines : a handbook of research in biomimetic and biohybrid systems . (Eds.) Tony J Prescott,Oxford, UK,Oxford University Press (2018),380-389 Ch. 39

Martin Weigert, Eugene W Myers, Moritz Kreysing
Biobeam - Rigorous wave-optical simulations of light-sheet microscopy
arXiv, Art. No. arXiv:1706.02261 (2017)
Open Access PDF

Sara N. Nagelberg, Lauren D. Zarzar, Natalie Nicolas, Kaushikaram Subramanian, Julia A Kalow, Vishnu Sresht, Daniel Blankschtein, George Barbastathis, Moritz Kreysing, Timothy M Swager, Mathias Kolle
Reconfigurable and responsive droplet-based compound micro-lenses.
Nat Commun, 8 Art. No. 14673 (2017)
Open Access PDF DOI
Micro-scale optical components play a crucial role in imaging and display technology, biosensing, beam shaping, optical switching, wavefront-analysis, and device miniaturization. Herein, we demonstrate liquid compound micro-lenses with dynamically tunable focal lengths. We employ bi-phase emulsion droplets fabricated from immiscible hydrocarbon and fluorocarbon liquids to form responsive micro-lenses that can be reconfigured to focus or scatter light, form real or virtual images, and display variable focal lengths. Experimental demonstrations of dynamic refractive control are complemented by theoretical analysis and wave-optical modelling. Additionally, we provide evidence of the micro-lenses' functionality for two potential applications-integral micro-scale imaging devices and light field display technology-thereby demonstrating both the fundamental characteristics and the promising opportunities for fluid-based dynamic refractive micro-scale compound lenses.

Mathias Kolle, Joseph D. Sandt, Sara N. Nagelberg, Lauren D. Zarzar, Moritz Kreysing, Peter Vukusic
Biological inspiration in optics and photonics: harnessing nature's light manipulation strategies for multifunctional optical materials (Invited talk)
In: Conference on Biophysics, Biology, and Biophotonics - The Crossroads (2016) Proceedings of SPIE; 9719, Bellingham, USA, SPIE (2016), 971903-971903

Moritz Kreysing, Lorenz Keil, Simon Lanzmich, Dieter Braun
Heat flux across an open pore enables the continuous replication and selection of oligonucleotides towards increasing length.
Nat Chem, 7(3) 203-208 (2015)
The replication of nucleic acids is central to the origin of life. On the early Earth, suitable non-equilibrium boundary conditions would have been required to surmount the effects of thermodynamic equilibrium such as the dilution and degradation of oligonucleotides. One particularly intractable experimental finding is that short genetic polymers replicate faster and outcompete longer ones, which leads to ever shorter sequences and the loss of genetic information. Here we show that a heat flux across an open pore in submerged rock concentrates replicating oligonucleotides from a constant feeding flow and selects for longer strands. Our experiments utilize the interplay of molecular thermophoresis and laminar convection, the latter driving strand separation and exponential replication. Strands of 75 nucleotides survive whereas strands half as long die out, which inverts the above dilemma of the survival of the shortest. The combined feeding, thermal cycling and positive length selection opens the door for a stable molecular evolution in the long-term microhabitat of heated porous rock.

Zuzanna Błaszczak, Moritz Kreysing, Jochen Guck
Direct observation of light focusing by single photoreceptor cell nuclei
Opt express, 22(9) 11043-11060 (2014)
Open Access PDF DOI

Mike Francke, Moritz Kreysing, Andreas Mack, Jacob Engelmann, Anett Karl, Felix Makarov, Jochen Guck, Mathias Kolle, Hartwig Wolburg, Roland Pusch, Gerhard von der Emde, Stefan Schuster, Hans-Joachim Wagner, Andreas Reichenbach
Grouped retinae and tapetal cups in some Teleostian fish: Occurrence, structure, and function.
Prog Retin Eye Res, 38 43-69 (2014)
This article presents a summary and critical review of what is known about the 'grouped retina', a peculiar type of retinal organization in fish in which groups of photoreceptor cell inner and outer segments are arranged in spatially separated bundles. In most but not all cases, these bundles are embedded in light-reflective cups that are formed by the retinal pigment epithelial cells. These cups constitute a specialized type of retinal tapetum (i.e., they are biological 'mirrors' that cause eye shine) and appear to be optimized for different purposes in different fishes. Generally, the large retinal pigment epithelial cells are filled with light-reflecting photonic crystals that consist of guanine, uric acid, or pteridine depending on species, and which ensure that the incoming light becomes directed onto the photoreceptor outer segments. This structural specialization has so far been found in representatives of 17 fish families; of note, not all members of a given family must possess a grouped retina, and the 17 families are not all closely related to each other. In many cases (e.g., in Osteoglossomorpha and Aulopiformes) the inner surface of the cup is formed by three to four layers of strikingly regularly shaped and spaced guanine platelets acting as an optical multilayer. It has been estimated that this provides an up to 10fold increase of the incident light intensity. In certain deep-sea fish (many Aulopiformes and the Polymixidae), small groups of rods are embedded in such 'parabolic mirrors'; most likely, this is an adaptation to the extremely low light intensities available in their habitat. Some of these fishes additionally possess similar tapetal cups that surround individual cones and, very likely, also serve as amplifiers of the weak incident light. In the Osteoglossomorpha, however, that inhabit the turbid water of rivers or streams, the structure of the cups is more complex and undergoes adaptation-dependent changes. At dim daylight, probably representing the usual environmental conditions of the fish, the outer segments of up to 30 cone cells are placed at the bottom of the cup where light intensity is maximized. Strikingly, however, a large number of rod receptor cells are positioned behind each mirroring cup. This peculiar arrangement (i) allows vision at deep red wavelenghts, (ii) matches the sensitivity of rod and cone photoreceptors, and (iii) facilitates the detection of low-contrast and color-mixed stimuli, within the dim, turbid habitat. Thus, for these fish the grouped retina appears to aid in reliable and quick detection of large, fast moving, biologically relevant stimuli such as predators. Overall, the grouped retina appears as a peculiar type of general retinal specialization in a variety of fish species that is adaptive in particular habitats such as turbid freshwater but also the deep-sea. The authors were prompted to write this review by working on the retina of Gnathonemus petersii; the data resulting from this work (Landsberger et al., 2008; Kreying et al., 2012) are included in the present review.

Moritz Kreysing, Dino Ott, Michael J Schmidberger, Oliver Otto, Markus Schürmann, Estela Martín-Badosa, Graeme Whyte, Jochen Guck
Dynamic operation of optical fibres beyond the single-mode regime facilitates the orientation of biological cells.
Nat Commun, 5 Art. No. 5481 (2014)
Open Access PDF DOI
The classical purpose of optical fibres is delivery of either optical power, as for welding, or temporal information, as for telecommunication. Maximum performance in both cases is provided by the use of single-mode optical fibres. However, transmitting spatial information, which necessitates higher-order modes, is difficult because their dispersion relation leads to dephasing and a deterioration of the intensity distribution with propagation distance. Here we consciously exploit the fundamental cause of the beam deterioration-the dispersion relation of the underlying vectorial electromagnetic modes-by their selective excitation using adaptive optics. This allows us to produce output beams of high modal purity, which are well defined in three dimensions. The output beam distribution is even robust against significant bending of the fibre. The utility of this approach is exemplified by the controlled rotational manipulation of live cells in a dual-beam fibre-optical trap integrated into a modular lab-on-chip system.

Mathias Kolle, Alfred Lethbridge, Moritz Kreysing, Jeremy J Baumberg, Joanna Aizenberg, Peter Vukusic
Bio-inspired band-gap tunable elastic optical multilayer fibers.
Adv Mater, 25(15) 2239-2245 (2013)
The concentrically-layered photonic structure found in the tropical fruit Margaritaria nobilis serves as inspiration for photonic fibers with mechanically tunable band-gap. The fibers show the spectral filtering capabilities of a planar Bragg stack while the microscopic curvature decreases the strong directional chromaticity associated with flat multilayers. Elongation of the elastic fibers results in a shift of the reflection of over 200 nm.

Moritz Kreysing, Roland Pusch, Dorothee Haverkate, Meik Landsberger, Jacob Engelmann, Janina Ruiter, Carlos Mora-Ferrer, Elke Ulbricht, Jens Grosche, Kristian Franze, Stefan Streif, Sarah Schumacher, Felix Makarov, Johannes Kacza, Jochen Guck, Hartwig Wolburg, James K Bowmaker, Gerhard von der Emde, Stefan Schuster, Hans-Joachim Wagner, Andreas Reichenbach, Mike Francke
Photonic crystal light collectors in fish retina improve vision in turbid water.
Science, 336(6089) 1700-1703 (2012)
Despite their diversity, vertebrate retinae are specialized to maximize either photon catch or visual acuity. Here, we describe a functional type that is optimized for neither purpose. In the retina of the elephantnose fish (Gnathonemus petersii), cone photoreceptors are grouped together within reflecting, photonic crystal-lined cups acting as macroreceptors, but rod photoreceptors are positioned behind these reflectors. This unusual arrangement matches rod and cone sensitivity for detecting color-mixed stimuli, whereas the photoreceptor grouping renders the fish insensitive to spatial noise; together, this enables more reliable flight reactions in the fish's dim and turbid habitat as compared with fish lacking this retinal specialization.

Moritz Kreysing
Optics of the retina: novel tools new insights
Ph.D. Thesis,University of Cambridge, Cambridge, United Kingdom (2011)

Moritz Kreysing, Lars Boyde, Jochen Guck, Kevin J Chalut
Physical insight into light scattering by photoreceptor cell nuclei.
Opt Lett, 35(15) 2639-2641 (2010)
A recent study showed that the rod photoreceptor cell nuclei in the retina of nocturnal and diurnal mammals differ considerably in architecture: the location of euchromatin and heterochromatin in the nucleus is interchanged. This inversion has significant implications for the refractive index distribution and the light scattering properties of the nucleus. Here, we extend previous two-dimensional analysis to three dimensions (3D) by using both a numerical finite-difference time-domain and an analytic Mie theory approach. We find that the specific arrangement of the chromatin phases in the nuclear core-shell models employed have little impact on the far-field scattering cross section. However, scattering in the near field, which is the relevant regime inside the retina, shows a significant difference between the two architectures. The "inverted" photoreceptor cell nuclei of nocturnal mammals act as collection lenses, with the lensing effect being much more pronounced in 3D than in two dimensions. This lensing helps to deliver light efficiently to the light-sensing outer segments of the rod photoreceptor cells and thereby improve night vision.

Irina Solovei, Moritz Kreysing, Christian Lanctôt, Süleyman Kösem, Leo Peichl, Thomas Cremer, Jochen Guck, Boris Joffe
Nuclear architecture of rod photoreceptor cells adapts to vision in mammalian evolution.
Cell, 137(2) 356-368 (2009)
We show that the nuclear architecture of rod photoreceptor cells differs fundamentally in nocturnal and diurnal mammals. The rods of diurnal retinas possess the conventional architecture found in nearly all eukaryotic cells, with most heterochromatin situated at the nuclear periphery and euchromatin residing toward the nuclear interior. The rods of nocturnal retinas have a unique inverted pattern, where heterochromatin localizes in the nuclear center, whereas euchromatin, as well as nascent transcripts and splicing machinery, line the nuclear border. The inverted pattern forms by remodeling of the conventional one during terminal differentiation of rods. The inverted rod nuclei act as collecting lenses, and computer simulations indicate that columns of such nuclei channel light efficiently toward the light-sensing rod outer segments. Comparison of the two patterns suggests that the conventional architecture prevails in eukaryotic nuclei because it results in more flexible chromosome arrangements, facilitating positional regulation of nuclear functions.

Moritz Kreysing, Tobias Kiessling, Anatol Fritsch, Christian Dietrich, Jochen Guck, Josef A Käs
The optical cell rotator.
Opt express, 16(21) 16984-16992 (2008)
Open Access PDF
The optical cell rotator (OCR) is a modified dual-beam laser trap for the holding and controlled rotation of suspended dielectric microparticles, such as cells. In contrast to optical tweezers, OCR uses two counter-propagating divergent laser beams, which are shaped and delivered by optical fibers. The rotation of a trapped specimen is carried out by the rotation of a dual-mode fiber, emitting an asymmetric laser beam. Experiments were performed on human erythrocytes, promyelocytic leukemia cells (HL60), and cell clusters (MCF-7). Since OCR permits the rotation of cells around an axis perpendicular to the optical axis of any microscope and is fully decoupled from imaging optics, it could be a suitable and expedient tool for tomographic microscopy.