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The healing of neuronal injuries is still an unachieved goal. Medicine-based therapies can only extend the survival of patients, but not finally lead to a healing process. Currently, a variety of stem cell-based tissue engineering developments are the subject of many research projects to bridge this gap. As yet, neuronal differentiation of induced pluripotent stem cells (iPS), embryonic cell lines, or neuronal stem cells could be accomplished and produce functional neuronally differentiated cells. However, clinical application of cells from these sources is hampered by ethical considerations. To overcome these hurdles numerous studies investigated the potential of adult mesenchymal stem cells (MSCs) as a potential stem cell source. Adult MSCs have been approved as cellular therapeutical products due to their regenerative potential and immunomodulatory properties. Only a few of these studies could demonstrate the capacity to differentiate MSCs into active firing neuron like cells. With this study we investigated the potential of Wharton’s Jelly (WJ) derived stem cells and focused on the intrinsic pluripotent stem cell pool and their potential to differentiate into active neurons. With a comprehensive neuronal differentiation protocol comprised of mechanical and biochemical inductive cues, we investigated the capacity of spontaneously forming stem cell spheroids (SCS) from cultured WJ stromal cells in regard to their neuronal differentiation potential and compared them to undifferentiated spheroids or adherent MSCs. Spontaneously formed SCSs show pluripotent and neuroectodermal lineage markers, meeting the pre-condition for neuronal differentiation and contain a higher amount of cells which can be differentiated into cells whose functional phenotypes in calcium and voltage responsive electrical activity are similar to neurons. In conclusion we show that up-concentration of stem cells from WJ with pluripotent characteristics is a tool to generate neuronal cell replacement. Graphical Abstract

ACS applied materials & interfaces 13(20), 23438-23451 (2021). doi:10.1021/acsami.1c03537, Published by American Chemical Society, Washington, DC

In this work, we analyze the impact of a chip coating with a self-assembled monolayer (SAM) of (3-aminopropyl)triethoxysilane (APTES) on the electronic and mechanical properties of neuroelectronic interfaces. We show that the large signal transfer, which has been observed for these interfaces, is most likely a consequence of the strong mechanical coupling between cells and substrate. On the one hand, we demonstrate that the impedance of the interface between Pt electrodes and an electrolyte is slightly reduced by the APTES SAM. However, this reduction of approximately 13% is definitely not sufficient to explain the large signal transfer of APTES coated electrodes demonstrated previously. On the other hand, the APTES coating leads to a stronger mechanical clamping of the cells, which is visible in microscopic images of the cell development of APTES-coated substrates. This stronger mechanical interaction is most likely caused by the positively charged amino functional group of the APTES SAM. It seems to lead to a smaller cleft between substrate and cells and, thus, to reduced losses of the cell's action potential signal at the electrode. The disadvantage of this tight binding of the cells to the rigid, planar substrate seems to be the short lifetime of the cells. In our case the density of living cells starts to decrease together with the visual deformation of the cells typically at DIV 9. Solutions for this problem might be the use of soft substrates and/or the replacement of the short APTES molecules with larger molecules or molecular multilayers.

An artificial lipid bilayer is the closest possible model for the cell membrane. Despite that, current methods of lipid bilayer assembly and functionalization do not provide a satisfactory mimic of the cell-cell contact due to the inability to recreate an asymmetrical multicomponent system. In the current work, a method to produce an integrated solid-supported lipid bilayer combining natural extracts from cell membranes and artificially made lipid vesicles is proposed. This simple method allows delivery of transmembrane proteins and components of the extracellular matrix into the substrate. Biocompatibility of the composite natural/artificial lipid bilayers is evaluated by their interactions with the cardiomyocyte-like HL-1 cell line. Compared with fully artificial mixes, composite lipid bilayers allow cells to adhere and develop a morphologically more normal cytoskeleton.

Small : nano micro 2200053 (2022). doi:10.1002/smll.202200053, Published by Wiley-VCH, Weinheim

Advanced biology 5(2), 2000248 (2021). doi:10.1002/adbi.202000248, Published by Wiley-VCH, Weinheim

In this paper, we demonstrate that cell adhesion and neuron maturation can be guided by patterned oxide surfaces functionalized with organic molecular layers. It is shown that the difference in the surface potential of various oxides (SiO2, Ta2O5, TiO2, and Al2O3) can be increased by functionalization with a silane, (3-aminopropyl)-triethoxysilane (APTES), which is deposited from the gas phase on the oxide. Furthermore, it seems that only physisorbed layers (no chemical binding) can be achieved for some oxides (Ta2O5 and TiO2), whereas self-assembled monolayers (SAM) form on other oxides (SiO2 and Al2O3). This does not only alter the surface potential but also affects the neuronal cell growth. The already high cell density on SiO2 is increased further by the chemically bound APTES SAM, whereas the already low cell density on Ta2O5 is even further reduced by the physisorbed APTES layer. As a result, the cell density is ∼8 times greater on SiO2 compared to Ta2O5, both coated with APTES. Furthermore, neurons form the typical networks on SiO2, whereas they tend to cluster to form neurospheres on Ta2O5. Using lithographically patterned Ta2O5 layers on SiO2 substrates functionalized with APTES, the guided growth can be transferred to complex patterns. Cell cultures and molecular layers can easily be removed, and the cell experiment can be repeated after functionalization of the patterned oxide surface with APTES. Thus, the combination of APTES-functionalized patterned oxides might offer a promising way of achieving guided neuronal growth on robust and reusable substrates.

Microelectrode arrays are widely used in different fields such as neurobiology or biomedicine to read out electrical signals from cells or biomolecules. One way to improve microelectrode applications is the development of novel electrode materials with enhanced or additional functionality. In this study, we fabricated macroelectrodes and microelectrode arrays containing gold penetrated by nanohole arrays as a conductive layer. We used this holey gold to optically excite surface plasmon polaritons, which lead to a strong increase in transparency, an effect that is further enhanced by the plasmon's interaction with cell culture medium. By varying the nanohole diameter in finite-difference time domain simulations, we demonstrate that the transmission can be increased to above 70% with its peak at a wavelength depending on the holey gold's lattice constant. Further, we demonstrate that the novel transparent microelectrode arrays are as suitable for recording cellular electrical activity as standard devices. Moreover, we prove using spectral measurements and finite-difference time domain simulations that plasmonically induced transmission peaks of holey gold red-shift upon sensing medium or cells in close vicinity (

The interface between cells and inorganic surfaces represents one of the key elements for bioelectronics experiments and applications ranging from cell cultures and bioelectronics devices to medical implants. In the present paper, we describe a way to tailor the biocompatibility of substrates in terms of cell growth and to significantly improve cell-chip communication, and we also demonstrate the reusability of the substrates for cell experiments. All these improvements are achieved by coating the substrates or chips with a self-assembled monolayer (SAM) consisting of a mixture of organic molecules, (3-aminopropyl)-triethoxysilane and (3-glycidyloxypropyl)-trimethoxysilane. By varying the ratio of these molecules, we are able to tune the cell density and live/dead ratios of rat cortical neurons cultured directly on the mixed SAM as well as neurons cultured on protein-coated SAMs. Furthermore, the use of the SAM leads to a significant improvement in cell-chip communications. Action potential signals of up to 9.4 ± 0.6 mV (signal-to-noise ratio up to 47) are obtained for HL-1 cells on microelectrode arrays. Finally, we demonstrate that the SAMs facilitate a reusability of the samples for all cell experiments with little re-processing.

Graphene based devices have already proven to be extremely sensitive and very useful in a wide spectrum of bioelectronics research. In the manuscript we describe a method to fabricate arrays of graphene-based probes, requiring minimal number of fabrication steps, while maintaining overall device functionality. These polyimide-based probes are approximately 6 μm thick, therefore ultraflexible, yet robust and stable. Devices, such as graphene field effect transistors (GFETs) and graphene multielectrode arrays (GMEAs) have been designed, fabricated and tested for their performance. The flexible GFETs exhibit sensitivity, i.e. transconductance up to 700 μS/V, which an order of magnitude larger compared to typical silicon transistors. Multiple probe per wafer design allows us to fabricate different kinds of devices on one 4-inch wafer, consequently increasing a possible range of applications from e.g. retinal to cortical neuroprosthetics.

Modifying the surfaces of oxides using self-assembled monolayers offers an exciting possibility to tailor their surface properties for various applications ranging from organic electronics to bioelectronics applications. The simultaneous use of different molecules in particular can extend this approach because the surface properties can be tuned via the ratio of the chosen molecules. This requires the composition and quality of the monolayers to be controlled on an organic level, that is, on the nanoscale. In this paper, we present a method of modifying the surface and surface properties of silicon oxide by growing self-assembled monolayers comprising various compositions of two different molecules, (3-aminopropyl)-triethoxysilane and (3-glycidyloxypropyl)-trimethoxysilane, by means of in situ controlled gas-phase deposition. The properties of the resulting mixed molecular monolayers (e.g., effective thickness, hydrophobicity, and surface potential) exhibit a perfect linear dependence on the composition of the molecular layer. Finally, coating the mixed layer with poly(l-lysine) proves that the density of proteins can be controlled by the composition as well. This indicates that the method might be an ideal way to optimize inorganic surfaces for bioelectronics applications.

Here, we present a method to investigate cell adhesion on soft, non-conducting polymers that are implant candidate materials. Neuronal cells were grown on two elastomers (polydimethylsiloxane (PDMS) and Ecoflex®) and prepared for electron microscopy. The samples were treated with osmium tetroxide (OsO4) and uranylacetate (UrAc). Best results can be achieved when the polymers were coated with a thin iridium layer before the cell culture. This was done to emphasize the usage of soft polymers as supports for implant electrodes. A good contrast and the adhesion of the cells on soft polymers could be visualized.

Magnetic materials have been widely used for various electromagnetic actuators. However, there still exists a corrosion problem for most magnetic materials when used in a biological cell environment. The metal ions released can cause cytotoxicity. In this paper, we characterize the corrosion of a soft-magnetic FeCo alloy, which has been adopted in our magnetic tweezers, in several cell culture media. Obvious accelerated corrosion was observed in cardiomyocyte and neuronal cell media, but not in Dulbecco’s Modified Eagle Medium. To eliminate the corrosion we examined electrochemical deposition of polypyrrole as well as pyrolytical deposition of parylene C for passivation. It was found that the quality of polypyrrole deposition was insufficient in the area near the edges of the tweezers tips where they had been laser-cut. However, the parylene C coating exhibited excellent isolation properties. In addition, parylene C-coated magnetic parts can also withstand repeated high magnetic field application. The cell viability of in vitro cultures exposed to the passivated material was inspected by live/dead cell staining. Both cardiomyocyte and primary neuronal cells could be cultured on parylene C coated magnetic parts for up to three weeks.

Organic electrochemical transistors (OECTs) have emerged as versatile electrophysiological sensors due to their high transconductance, biocompatibility, and transparent channel material. High maximum transconductances are demonstrated facilitating the extracellular recording of signals from electrogenic cells. However, this requires large channel dimensions and thick polymer films. These large channel dimensions lead to low transistor densities. Here, interdigitated OECTs (iOECTs) are introduced, which feature high transconductances at small device areas. A superior device performance is achieved by systematically optimizing the electrode layout regarding channel length, number of electrode fingers and electrode width. Interestingly, the maximum transconductance (g(max)) does not straightforwardly scale with the channel width-to-length ratio, which is different from planar OECTs. This deviation is caused by the dominating influence of the source-drain series resistance R-sd for short channel devices. Of note, there is a critical channel length (15 mu m) above which the channel resistance R-ch becomes dominant and the device characteristics converge toward those of planar OECTs. Design rules for engineering the performance of iOECTs are proposed and tested by recording action potentials of cardiomyocyte-like HL-1 cells with high signal-to-noise ratios. These results demonstrate that interdigitated OECTs meet two requirements of bioelectronic applications, namely, high device performance and small channel dimensions.

In recent years, optogenetic approaches have significantly advanced the experimental repertoire of cellular and functional neuroscience. Yet, precise and reliable methods for specific expression of optogenetic tools remain challenging. In this work, we studied the transduction efficiency of seven different adeno-associated virus (AAV) serotypes in primary cortical neurons and revealed recombinant (r) AAV6 to be the most efficient for constructs under control of the cytomegalovirus (CMV) promoter. To further specify expression of the transgene, we exchanged the CMV promoter for the human synapsin (hSyn) promoter. In primary cortical-glial mixed cultures transduced with hSyn promoter-containing rAAVs, expression of ChR2opt (a Channelrhodopsin-2 variant) was limited to neurons. In these neurons action potentials could be reliably elicited upon laser stimulation (473nm). The use of rAAV serotype alone to restrict expression to neurons results in a lower transduction efficiency than the use of a broader transducing serotype with specificity conferred via a restrictive promoter. Cells transduced with the hSyn driven gene expression were able to elicit action potentials with more spatially and temporally accurate illumination than neurons electrofected with the CMV driven construct. The hSyn promoter is particularly suited to use in AAVs due to its small size. These results demonstrate that rAAVs are versatile tools to mediate specific and efficient transduction as well as functional and stable expression of transgenes in primary cortical neurons.

The present study of nanoelectrochemical sensors prepared by directed electrochemical nanowire assembly (DENA) is defined by the requirements of electrochemical analysis, where the transducer function of metallic nanowires is synergetically combined with their electrochemical catalytic activity with respect to a particular analyte. We show for the first time that this technique can be employed for metals (Pd, Au) and their bimetallic compositions to create various multicomponent sensor nanomaterials on a single chip without the use of multistep lithography for the spatially resolved analysis of solutions. The nanostructures of various compositions can be individually addressed when used in liquid media, so that the particular surface properties of the individual nanoarray elements can be used for the electrochemical analysis of specific analytes. The sensor application of these devices in electrolytes and cell culture conditions has been demonstrated for the first time. As an example, the Pd-Au nanowires prepared by DENA were used for a non-enzymatic analysis of H2O2 with a linear concentration interval of 10−6–10−3 M, sensitivity of 18 μA M−1, and detection limit of 3 × 10−7 M at as low absolute value of the detection potential as − 0.05 V. This sensor was also proven for the detection of hydrogen peroxide in HL-1 cell culture, demonstrating good biocompatibility and support for the cell culture conditions. Using various DENA-grown electrochemical compositions on a single chip, a novel multisensor platform is proposed for the determination of various analytes in electrolyte solutions for biocompatible sensor arrays, flexible multianalyte environmental and technological process monitoring, and healthcare areas.

ChR2-XXL and GtACR1 are currently the cation and anion ends of the optogenetic single channel current range. These were used in primary rat cortical neuronsin vitroto manipulate neuronal firing patterns. ChR2-XXL provides high cation currents via elevated light sensitivity and a prolonged open state. Stimulating ChR2-XXL expressing putative presynaptic neurons induced neurotransmission. Moreover, stable depolarisation block could be generated in single neurons using ChR2-XXL, proving that ChR2-XXL is a promising candidate forin vivoapplications of optogenetics, for example to treat peripheral neuropathic pain. We also addressed an anion channelrhodopsin (GtACR1) for the next generation of optogenetic neuronal inhibition in primary rat cortical neurons. GtACR1‘s light-gated chloride conduction was verified in primary neurons and the efficient photoinhibition of action potentials, including spontaneous activity, was shown. Our data also implies that the chloride concentration in neurons decreases during neural development. In both cases, we find surprising applications of these high current channels. For ChR2-XXL inhibition and stimulation are possible, while for GtACR1 the role of Cl−during neural development becomes a new optogenetic target.

International audience; A recently discovered material, carbon nanotubes covered with diamond (DCNTs) was tested for its suitability in bioelectronics applications. Diamond shows advantages for bioelectronics applications (wide electro chemical window and bioinertness). This study investigates the effect of electrode surface shape (flat or three dimensional) on cell growth and behavior. For comparison, flat nanocrystalline diamond substrates were used. Primary embryonic neurons were grown on top of the structures and neither incorporated the structures nor did they grow in between the single structures. The interface was closely examined using focused ion beam (FIB) and scanning electron microscopy. Of special interest was the interface between cell and substrate. 5% to 25% of the cell membrane adhered to the substrate, which fits the theoretical estimated value. While investigating the conformity of the neurons, it could be observed that the cell membrane attaches to different heights of the tips of the 3D structure. However, the aspect ratio of the structures had no effect on the cell viability. These results let us assume that not more than 25% of cell attachment is needed for the survival of a functional neuronal cell.

International audience; The expansion of diamond-based electronics in the area of biological interfacing has not been as thoroughly explored as applications in electrochemical sensing. However, the biocompatibility of diamond, large safe electrochemical window, stability, and tunable electronic properties provide opportunities to develop new devices for interfacing with electrogenic cells. Here, the fabrication of microelectrode arrays (MEAs) with boron-doped nanocrystalline diamond (BNCD) electrodes and their interfacing with cardiomyocyte-like HL-1 cells to detect cardiac action potentials are presented. A non-reductive means of structuring doped and undoped diamond on the same substrate is shown. The resulting BNCD electrodes show high stability under mechanical stress generated by the cells. It is shown that by fabricating the entire surface of the MEA with NCD, in patterns of conductive doped, and isolating undoped regions, signal detection may be improved up to four-fold over BNCD electrodes passivated with traditional isolators.

There is increasing evidence that mechanical issues play a vital role in neuron growth and brain development. The importance of this grows as novel devices, whose material properties differ from cells, are increasingly implanted in the body. In this work, we studied the mechanical properties of rat brain cells over time and on different materials by using a high throughput magnetic tweezers system. It was found that the elastic moduli of both neurite and soma in networked neurons increased with growth. However, neurites at DIV4 exhibited a relatively high stiffness, which could be ascribed to the high outgrowth tension. The power-law exponents (viscoelasticity) of both neurites and somas of neurons decreased with culture time. On the other hand, the stiffness of glial cells also increased with maturity. Furthermore, both neurites and glia become softer when cultured on compliant substrates. Especially, the glial cells cultured on a soft substrate obviously showed a less dense and more porous actin and GFAP mesh. In addition, the viscoelasticity of both neurites and glia did not show a significant dependence on the substrates' stiffness.

Bioactive molecules such as adhesion ligands, growth factors, or enzymes play an important role in modulating cell behavior such as cell adhesion, spreading, and differentiation. Deciphering the mechanism of ligand-mediated cell adhesion and associated signaling is of great interest not only for fundamental biophysical investigations but also for applications in medicine and biotechnology. In the presented work, we developed a new biomimetic platform that enables culturing primary neurons and testing cell surface-receptor ligand interactions in cell-cell contacts as, e.g., in neuronal synapses. This platform consists of a supported lipid bilayer modified with incorporated neuronal adhesion proteins conjugated with the Fc-domain of IgG (ephrin A5 Fc-chimera). We extensively characterized properties of these protein containing bilayers using fluorescence recovery after photobleaching (FRAP), quartz crystal microbalance with dissipation (QCM-D), and immunostaining. We conclude that the Fc-domain is the part responsible for the incorporation of the protein into the bilayer. The biomimetic platform prepared by this new approach was able to promote neuronal cell adhesion and maintain growth as well as facilitate neuronal maturation as shown by electrophysiological measurements. We believe that our approach can be extended to insert other proteins to create a general culture platform for neurons and other cell types.

Flexible and transparent electronic devices possess crucial advantages over conventional silicon based systems for bioelectronic applications since they are able to adapt to nonplanar surfaces, cause less chronic immunoreactivity, and facilitate easy optical inspection. Here, organic electrochemical transistors (OECTs) are embedded in a flexible matrix of polyimide to record cardiac action potentials. The wafer-scale fabricated devices exhibit transconductances (12 mS V-1 ) and drain-source on-to-off current ratios (≈105 ) comparable to state of the art nonflexible and superior to other reported flexible OECTs. The transfer characteristics of the devices are preserved even after experiencing extremely high bending strain and harsh crumpling. A sub-micrometer poly(3,4-ethylenedioxythiophene) doped with poly(styrenesulfonate) layer results in a fast transport of ions between the electrolyte and the polymer channel characterized by a cut-off frequency of 1200 Hz. Excellent device performance is proved by mapping the propagation of cardiac action potentials with high signal-to-noise ratio. These results demonstrate that the electrical performance of flexible OECTs can compete with hard-material-based OECTs and thus potentially be used for in vivo applications.

An analysis of 105 mergers among small UK biotech companies over a 10-year period shows that the improvement of shareholder positions, rather than product pipelines or business opportunities, is the main motivation for such transactions.

Graphene multielectrode arrays (GMEAs) presented in this work are used for cardio and neuronal extracellular recordings. The advantages of the graphene as a part of the multielectrode arrays are numerous: from a general flexibility and biocompatibility to the unique electronic properties of graphene. The devices used for extensive in vitro studies of a cardiac-like cell line and cortical neuronal networks show excellent ability to extracellularly detect action potentials with signal to noise ratios in the range of 45 ± 22 for HL-1 cells and 48 ± 26 for spontaneous bursting/spiking neuronal activity. Complex neuronal bursting activity patterns as well as a variety of characteristic shapes of HL-1 action potentials are recorded with the GMEAs. This paper illustrates that the potential applications of the GMEAs in biological and medical research are still numerous and diverse.

Genetically encoded light-sensitive proteins can be used to manipulate and observe cellular functions. According to different modes of action, these proteins are divided into actuators like the blue-light gated cation channel Channelrhodopsin-2 (ChR2) and detectors like the calcium sensor GCaMP. In order to optogenetically control and study the activity of rat primary cortical neurons, we established a transduction procedure using recombinant Adeno-associated viruses (rAAVs) as gene-ferries. Thereby, we achieved high transduction rates of these neurons with ChR2. In ChR2 expressing neurons, action potentials could be repeatedly and precisely elicited with laser pulses and measured via patch clamp recording.

We present an optoelectrical device capable of in vitro optical stimulation and electrophysiological recording. The device consists of an array of micropixellated InGaN light-emitting diodes coupled to a custom-made ultrathin planar microelectrode array. Cells can be cultured directly on the chip for short- and long-term electrophysiological experiments. To show the functionality of the device, we transfected a cardiomyocyte-like cell line (HL-1) with a light-sensitive protein channelrhodopsin. We monitored action potentials of individual, spontaneously beating, HL-1 cells growing on the chip by extracellular electrical recordings. On-chip optical stimulation was demonstrated by triggering network activity in a confluent HL-1 cell culture and visualized by calcium imaging. We see the potential of our system for electrophysiological experiments with optogenetically modified cells. Optical stimulation can be performed directly on the chip without additional optical components or external light sources.

Recording extracellular potentials from electrogenic cells (especially neurons) is the hallmark destination of modern bioelectronics. While fabrication of flexible and biocompatible in vivo devices via silicon technology is complicated and time-consuming, graphene field-effect transistors (GFETs), instead, can easily be fabricated on flexible and biocompatible substrates. In this work, we compare GFETs fabricated on rigid (SiO2 /Si and sapphire) and flexible (polyimide) substrates. The GFETs, fabricated on the polyimide, exhibit extremely large transconductance values, up to 11 mS·V–1, and mobility over 1750 cm2 ·V–1·s–1. In vitro recordings from cardiomyocyte-like cell culture are performed by GFETs on a rigid transparent substrate (sapphire). Via multichannel measurement, we are able to record and analyze both: difference in action potentials as well as their spatial propagation over the chip. Furthermore, the controllably flexible polyimide-on-steel (PIonS) substrates are able to ex vivo record electrical signals from primary embryonic rat heart tissue. Considering the flexibility of PIonS chips, together with the excellent sensitivity, we open up a new road into graphene-based in vivo biosensing.

We present a new biocompatible nanostructured microelectrode array for extracellular signal recording from electrogenic cells. Microfabrication techniques were combined with a template-assisted approach using nanoporous aluminum oxide to develop gold nanopillar electrodes. The nanopillars were approximately 300-400 nm high and had a diameter of 60 nm. Thus, they yielded a higher surface area of the electrodes resulting in a decreased impedance compared to planar electrodes. The interaction between the large-scale gold nanopillar arrays and cardiac muscle cells (HL-1) was investigated via focused ion beam milling. In the resulting cross-sections we observed a tight coupling between the HL-1 cells and the gold nanostructures. However, the cell membranes did not bend into the cleft between adjacent nanopillars due to the high pillar density. We performed extracellular potential recordings from HL-1 cells with the nanostructured microelectrode arrays. The maximal amplitudes recorded with the nanopillar electrodes were up to 100% higher than those recorded with planar gold electrodes. Increasing the aspect ratio of the gold nanopillars and changing the geometrical layout can further enhance the signal quality in the future.

During development individual cells in tissues undergo complex cell-shape changes to drive the morphogenetic movements required to form tissues. Cell shape is determined by the cytoskeleton and cell-shape changes critically depend on a tight spatial and temporal control of cytoskeletal behavior. We have used the formation of the salivary glands in the Drosophila embryo, a process of tubulogenesis, as an assay for identifying factors that impinge on cell shape and the cytoskeleton. To this end we have performed a gain-of-function screen in the salivary glands, using a collection of fly lines carrying EP-element insertions that allow the overexpression of downstream-located genes using the UAS-Gal4 system. We used a salivary-gland-specific fork head-Gal4 line to restrict expression to the salivary glands, in combination with reporters of cell shape and the cytoskeleton. We identified a number of genes known to affect salivary gland formation, confirming the effectiveness of the screen. In addition, we found many genes not implicated previously in this process, some having known functions in other tissues. We report the initial characterization of a subset of genes, including chickadee, rhomboid1, egalitarian, bitesize, and capricious, through comparison of gain- and loss-of-function phenotypes.

We characterized the mechanical properties of cardiomyocyte-like HL-1 cells using our recently developed multi-pole magnetic tweezers. With the optimized design, both high force and high throughput are achieved at the same time. Force up to 100 pN can be applied on a 1 μm diameter superparamagnetic bead in a workspace with 60 μm radius, which is encircled symmetrically by 3 sharp magnetic tips. By adjusting the coil currents, both the strength and direction of force can be controlled. The result shows that both viscosity and shear elastic modulus of HL-1 cells exhibit an approximately log-normal distribution. The cells became stiffer as they matured, consistent with a transition from proliferating cells to contractile muscle tissue. Moreover, the mechanical properties of HL-1 cells show high heterogeneity, which agrees well with their physiological structure.

This article shows the combination of light activatable ion channels and microelectrode array (MEA) technology for bidirectionally interfacing cells. HL-1 cultures, a mouse derived cardiomyocyte-like cell line, transfected with channelrhodopsin were stimulated with a microscope coupled 473 nm laser and recorded with custom built 64 electrode MEAs. Channelrhodopsin induced depolarization of the cell can evoke action potentials (APs) in single cells. Spreading of the AP over the cell layer can then be measured with good spatiotemporal resolution using MEA recordings. The possibility for light induced pacemaker switching in cultures was shown. Furthermore, the suppression of APs can also be achieved with the laser. Possible applications include cell analysis, e.g. pacemaker interference or induced pacemaker switching, and medical applications such as a combined cardiac pacemaker and defibrillator triggered by light. Since current prosthesis research focuses on bidirectionality, this system may be applied to any electrogenic cell, including neurons or muscles, to advance this field.