3D-printed Holder Research Articles

A Pd/Al2O3-based micro-reformer unit fully integrated in silicon technology for H-rich gas production

This work reports the design, manufacturing and catalytic activity characterization of a micro-reformer for hydrogen-rich gas generation integrated in portable-solid oxide fuel cells (μ-SOFCs). The reformer has been designed as a silicon micro monolithic substrate compatible with the mainstream microelectronics fabrication technologies ensuring a cost-effective high reproducibility and reliability. Design and geometry of the system have been optimized comparing with the previous design, consisting in an array of more than 7x103 vertical through-silicon micro channels perfectly aligned (50 μm diameter) and a 5 W integrated serpentine heater consisting of three stacked metallic layers (TiW, W and Au) for perfect adhesion and passivation. Traditional fuels for SOFCs, such as ethanol or methanol, have been replaced by dimethyl ether (DME) and the chosen catalyst for DME conversion consists of Pd nanoparticles grafted on an alumina active support. The micro-channels have been coated by atomic layer deposition (ALD) with amorphous Al2O3 and the influence of rapid thermal processing (RTP) on such film has been studied. A customized ceramic 3D-printed holder has been designed to measure the specific hydrogen production rates, DME conversion and selectivity profiles of such catalyst at different temperatures.

Ptychographic modulation engine: a low-cost DIY microscope add-on for coherent super-resolution imaging

Imaging of biological cells and tissues often relies on fluorescent labels, which offer high contrast with molecular specificity. The use of exogenous labeling agents, however, may alter the normal physiology of the bio-specimens. Complementary to the established fluorescence microscopy, label-free quantitative phase imaging provides an objective morphological measurement tool for bio-specimens and is free of variability introduced by contrast agents. Here, we report a simple and low-cost microscope add-on, termed ptychographic modulation engine (PME), for super-resolution quantitative phase imaging. In this microscope add-on module, we attach a diffuser to a 3D-printed holder that can be mechanically moved to different x-y positions. We then use two vibrational motors to introduce random positional shifts to the diffuser. The add-on module can be placed between the objective lens and the specimen in most existing microscope platforms. Thanks to the diffuser modulation process, the otherwise inaccessible high-resolution object information can now be encoded into the captured images. In the ptychographic phase retrieval process, we jointly recover the complex object wavefront, the complex diffuser profile, and the unknown positional shifts of the diffuser. We demonstrate a four-fold resolution gain over the diffraction limit of the employed 2× objective lens. We also test our approach for in vitro cell imaging, where we are able to adjust the focus after the data has been captured. The reported microscope add-on provides a turnkey solution for super-resolution quantitative phase imaging. It may find applications in label-free bio-imaging where both large field-of-view and high resolution are needed.

Rapid-scan EPR imaging of a phantom comprised of species with different linewidths and relaxation times.

As a demonstration of the application of rapid-scan EPR to imaging at low frequency and magnetic field, a multi-compartment phantom containing six different samples was imaged. The samples were nitroxide radicals, trityl (substituted triarylmethyl) radicals, and the oxygen-sensitive solid lithium phthalocyanine (LiPc), all of which are useful for in vivo imaging. The 2D spectral-spatial image demonstration was performed at 250 MHz, with samples in sealed tubes of various sizes arranged in a 3D-printed plastic holder. Maximum gradients of 10 G/cm gave a spatial resolution of about 0.1 mm for the narrow trityl and LiPc signals and about 1 mm for the nitroxide. The importance of proper selection of resonator bandwidth and scan rate for obtaining accurate linewidth information is demonstrated for a case in which the phantom is composed of species with signal linewidths and relaxation times that differ by more than a factor of 10.

Low-cost (

Spectrophotometry is a fundamental technique in many areas of science, with many applications and uses. The cost of spectrophotometers has acted as a barrier on the teaching and use of the technique. Here, we provide open-source plans to a 3D-printed cuvette holder with an interchangeable narrow–spectral bandwidth light-emitting diode (LED) block that can be used in conjunction with a smartphone’s ambient light sensor (ALS) to perform spectrophotometry. A Lego version with an interchangeable LED block is also presented. Results from the smartphone spectrophotometer in comparison with commercially available spectrophotometers demonstrated functionality, and the model may have many applications, especially in indirect spectrophotometry, such as in the protein assay shown here. The plans for the 3D-printed model are freely available on GitHub, as are editable files to allow customisation by users. We would encourage users to share adaptations with the scientific community.

Salivary diagnostics on paper microfluidic devices and their use as wearable sensors for glucose monitoring.

Microfluidic paper-based devices (μPADs) and wearable devices have been highly studied to be used as diagnostic tools due to their advantages such as simplicity and ability to provide instrument-free fast results. Diseases such as periodontitis and diabetes mellitus can potentially be detected through these devices by the detection of important biomarkers. This study describes the development of μPADs through craft cutter printing for glucose and nitrite salivary diagnostics. In addition, the use of μPADs integrated into a mouthguard as a wearable sensor for glucose monitoring is also presented. μPADs were designed to contain two detection zones for glucose and nitrite assays and a sampling zone interconnected by microfluidic channels. Initially, the analytical performance of the proposed μPADs was investigated and it provided linear behavior (r2 ≥ 0.994) in the concentration ranges between 0 to 2.0mmolL-1 and 0 to 400μmolL-1 for glucose and nitrite, respectively. Under the optimized conditions, the limits of detection achieved for glucose and nitrite were 27μmolL-1 and 7μmolL-1, respectively. Human saliva samples were collected from healthy individuals and patients previously diagnosed with periodontitis or diabetes and then analyzed on the proposed μPADs. The results found using μPADs revealed higher glucose concentration values in saliva collected from patients diagnosed with diabetes mellitus and greater nitrite concentrations in saliva collected from patients diagnosed with periodontitis, as expected. The results obtained on μPADs did not differ statistically from those measured by spectrophotometry. With the aim of developing paper-based wearable sensors, μPADs were integrated, for the first time, into a silicone mouthguard using a 3D-printed holder. The proof of concept was successfully demonstrated through the monitoring of the glucose concentration in saliva after the ingestion of chocolate. According to the results reported herein, paper-based microfluidic devices offer great potential for salivary diagnostics, making their integration into a silicone mouthguard possible, generating simple, low-cost, instrument-free, and powerful wearable sensors.

Microfluidic oxygen sensor system as a tool to monitor the metabolism of mammalian cells

We present a sensor system to monitor the uptake of oxygen by mammalian cells as a direct indicator for the metabolism under consideration of the requirements for the usage in biolabs. That includes reliable oxygen sensing as well as a small footprint of the setup without sophisticated external equipment, the usage of compatible materials and the suitability for a high degree of integration and automation. These requirements are fulfilled by a system that consists of a microfluidic chip with an integrated oxygen-sensitive phosphorescent film, heater and temperature sensor, external optical read-out and 3D-printed holders and housing. The chip with the closed microfluidic chamber is made by clean-room technologies out of silicon and glass. An excitation LED and a small and low-cost Raspberry Pi camera are used to measure the phosphorescent signal. With this system, the concentration of dissolved oxygen can be determined with an accuracy of ±0.8% (air) for oxygen concentrations between 0 and 26% (air) and at any temperature between 23 and 41 °C. To demonstrate the capabilities, the oxygen consumption rates of HaCaT-cells (human keratinocyte cell line) are determined at different temperatures.

Direct Coupling of Dispersive Extractions with Magnetic Particles to Mass Spectrometry via Microfluidic Open Interface.

Microextraction coupled to mass spectrometry (MS) has great potential in analytical chemistry laboratories operating in a variety of fields. Indeed, microextraction methods directly coupled to MS can be of large value given that they can provide not only analyte extraction and enrichment but also effective sample cleanup. In recent years, the practicality in handling high active surface area, versatility, and environmentally friendly features of magnetic dispersive microextraction technologies has contributed to an explosion in the number of methods and technologies reported in the literature for a wide range of applications. However, to the best of our knowledge, no technology to date has been capable of efficiently merging these two rising concepts in a simple and integrated analytical workflow. In this context, the microfluidic open interface is presented for the direct coupling of dispersive magnetic extraction to mass spectrometry. This technology operates under the concept of a flow-isolated desorption volume, which generates a stagnant droplet open to ambient conditions while continuously feeding the ionization source with solvent by means of the self-aspiration process intrinsic of the electrospray ionization (ESI) interface. To improve the efficiency of the final analytical workflow, a novel dispersive magnetic micro- and nanoparticle extraction protocol for biofluid droplets was developed. The final methodology entailed the dispersion of a small amount of magnetic particles (20-70 μg) in a droplet of biofluid (≤40 μL) for extraction, followed by a particle collection step using a homemade 3D-printed holder containing an embedded rare-earth magnet. In the final step, the holder is set on top of the microfluidic open interface (MOI) for desorption in the isolated droplet. Switching the valve transfers the desorbed analytes to the ESI source in less than 5 s. As proof of concept, the completely new setup was applied to the determination of prohibited substances from phosphate-buffered saline (PBS) and human urine using Fe2O3 magnetic nanoparticles (50 nm) functionalized with C18. The limits of quantitation (LOQs) obtained were in the low-ppb range in all cases, and acceptable precision (≤20%) and accuracy (80-120%) were attained. Also, taking advantage of the fast extraction kinetics provided by the radial diffusion associated with small particles, we employed the methodology for the selective extraction of phosphopeptides from 40 μL of tryptic β-casein digest using 70 μg of magnetic Ti-IMAC microparticles. To conclude, the technology and methodology herein presented provided excellent capabilities comparable to those of other solid-phase microextraction (SPME-MS) approaches while dramatically minimizing the amount of sample and sorbent required per analysis, as well as affording significantly fast extraction times due to the enhanced kinetics of extraction.

Dissolvable Polyvinyl-Alcohol Film, a Time-Barrier to Modulate Sample Flow in a 3D-Printed Holder for Capillary Flow Paper Diagnostics.

Integrating a dissolvable membrane into a sensor allows the control of sample flow, location and duration in critical areas. These time-barrier films stop the flow of samples until the membrane has dissolved, thus, for example, allowing increased exposure time between immunoreagents for the formation of greater numbers of immuno-complexes, ensuring higher sensitivity, reactivity, and helping to reduce false-positive signals. In this study, dissolvable polyvinyl alcohol (PVA) films are used in a 3D-printed sensor holder, which enables film integration without the use of glue. PVA is a synthetic hydrophilic linear polymer, its solubility is dependent on its molecular weight and degree of hydrolysis. Three types of PVAs films were tested herein: (1) PVA 1-Mw: 30–70 K, 87–90% hydrolyzed; (2) PVA 2-Mw: 31–50 K, 98–99% hydrolyzed and (3) PVA 3-Mw: 89–98 K, >99% hydrolyzed. The films were exposed to water in (1) the novel 3D-printed holder and (2) directly immersed into a water droplet. After comparing the time taken to dissolve PVA 1–3 films, PVA 1 films of 5–20% (w/v) are found to be most suitable as time barrier films, due to their optimal dissolution times and physical properties for integration into the customized 3D-printed holder.

A miniaturized, tethered, spectrally-encoded confocal endomicroscopy capsule.

The tethered spectrally-encoded confocal endomicroscopy (SECM) capsule is an imaging device that once swallowed by an unsedated patient can visualize cellular morphologic changes associated with gastrointestinal (GI) tract diseases in vivo. Recently, we demonstrated a tethered SECM capsule for counting esophageal eosinophils in patients with eosinophilic esophagitis (EoE) in vivo. Yet, the current tethered SECM capsule is far too long to be widely utilized for imaging pediatric patients, who constitute a major portion of the EoE patient population. In this paper, we present a new tethered SECM capsule that is 33% shorter, has an easier and repeatable fabrication process, and produces images with reduced speckle noise. The smaller SECM capsule utilized a miniature condenser to increase the fiber numerical aperture and reduce the capsule length. A custom 3D-printed holder was developed to enable easy and repeatable device fabrication. A dual-clad fiber (DCF) was used to reduce speckle noise. The fabricated SECM capsule (length = 20 mm; diameter = 7 mm) had a similar size and shape to a pediatric dietary supplement pill. The new capsule achieved optical sectioning thickness of 13.2 μm with a small performance variation between devices of 1.7 μm. Confocal images of human esophagus obtained in vivo showed the capability of this new device to clearly resolve microstructural epithelial details with reduced speckle noise. We expect that the smaller size and better image performance of this new SECM capsule will greatly facilitate the clinical adoption of this technology in pediatric patients and will enable more accurate assessment of EoE-suspected tissues. Lasers Surg. Med. 51:452-458, 2019. © 2019 Wiley Periodicals, Inc.

3D-printing enabled micro-assembly of a microfluidic electroporation system for 3D tissue engineering.

Electro-transfection is an essential workhorse tool for regulating cellular responses and engineering cellular materials in tissue engineering. However, most of the existing approaches are only focused on cell suspensions in vitro, which fails to mimic an in vivo tissue microenvironment regarding the 3D electric field distribution and mass transport in a biological matrix. However, building a 3D electro-transfection system that is compatible with 3D cell culture for mimicking the in vivo tissue microenvironment is challenging, due to the substantial difficulties in control of the 3D electric field distribution as well as the cellular growth. To address such challenges, we introduce a novel 3D micro-assembly strategy assisted by 3D printing, which enables the molding of 3D microstructures as LEGO® parts from 3D-printed molds. The molded PDMS LEGO® bricks are then assembled into a 3D-cell culture chamber interconnected with vertical and horizontal perfusion microchannels as a 3D channel network. Such a 3D perfusion microchannel network is unattainable by direct 3D printing or other microfabrication approaches, which can facilitate the highly-efficient exchange of nutrition and waste for 3D cell growth. Four flat electrodes are mounted into the 3D culture chamber via a 3D-printed holder and controlled by a programmable power sequencer for multi-directional electric frequency scanning (3D μ-electro-transfection). This multi-directional scanning not only can create transient pores all over the cell membrane, but also can generate local oscillation for enhancing mass transport and improving cell transfection efficiency. As a proof-of-concept, we electro-delivered the pAcGFP1-C1 vector to 3D cultured HeLa cells within peptide hydrogel scaffolding. The expressed GFP level from transfected HeLa cells reflects the transfection efficiency. We found two key parameters including electric field strength and plasmid concentration playing more important roles than the pulse duration and duty cycles. The results showed an effective transfection efficiency of ∼15% with ∼85% cell viability, which is 3-fold higher compared to that of the conventional benchtop 3D cell electro-transfection. This 3D μ-electrotransfection system was further used for genetically editing 3D-cultured Hek-293 cells via direct delivery of CRISPR/Cas9 plasmid which showed successful transfection with GFP expressed in the cytoplasm as the reporter. The 3D-printing enabled micro-assembly allows facile creation of a novel 3D culture system for electro-transfection, which can be employed for versatile gene delivery and cellular engineering, as well as building in vivo like tissue models for fundamentally studying cellular regulation mechanisms at the molecular level.

A smartphone-based system for fluorescence polarization assays

This paper demonstrates the use of a smartphone-based sensor for fluorescence polarization (FP) analysis of biomolecules. The FP detection can rapidly sense ligand-analyte bindings by measuring molecule mobility, and thus, FP-based assays have been widely used for rapid diagnostics in clinics. Here, we implemented the FP detection apparatus using a 3D-printed compact holder and the built-in camera of a smartphone. The system offers accurate measurements of the degree of polarization by simultaneously detecting the fluorescence intensities parallel and perpendicular to the polarization of the excitation. The fluorescence signal of the sample is excited by a laser or light-emitting diode and separated by a polarization beam cube depending on the polarization. Parallel and perpendicular polarized emissions are projected onto two different regions of the sensor chip in the smartphone camera. A custom software app was developed to count the average intensity in the areas of interest and compute the degree of polarization. We validated the system by measuring the polarization of dye molecules dissolved in solutions with different viscosities. As an example of biomolecule sensing, a competitive FP immunoassay of Prostaglandin E2 was demonstrated using the developed system and exhibited the limit of detection of 1.57 ng/mL. The smartphone-based FP assay platform can also be implemented for the detection of toxins, disease biomarkers, and pathogens in resource-limited settings.

First measurements of a scintillator based fast-ion loss detector near the ASDEX Upgrade divertor.

A new reciprocating scintillator based fast-ion loss detector has been installed a few centimeters above the outer divertor of the ASDEX Upgrade tokamak and between two of its lower Edge Localized Modes (ELM) mitigation coils. The detector head containing the scintillator screen, Faraday cup, calibration lamp, and collimator systems are installed on a motorized reciprocating system that can adjust its position via remote control in between plasma discharges. Orbit simulations are used to optimize the detector geometry and velocity-space coverage. The scintillator image is transferred to the light acquisition systems outside of the vacuum via a lens relay (embedded in a 3D-printed titanium holder) and an in-vacuum image guide. A charge coupled device camera, for high velocity-space resolution, and an 8 × 8 channel avalanche photo diode camera, for high temporal resolution (up to 2 MHz), are used as light acquisition systems. Initial results showing velocity-space of neutral beam injection prompt losses and fast-ion losses induced by a (2, 1) neoclassical tearing mode are presented.

Influence of stacking fault energy and dislocation character on slip transfer at coherent twin boundaries studied by micropillar compression

Copper and α-brass micropillars containing a single coherent twin boundary of controlled orientation were compressed to systematically investigate dislocation-twin boundary interactions as a function of the dislocation type and generalized stacking fault energies. For this purpose, bicrystalline micropillars in [112], [110] and [259] orientations were prepared from polycrystalline samples using focused ion beam milling in combination with custom 3D-printed sample holders. Consistent with previous findings, a vertical twin boundary neither resulted in the storage of dislocations at the interface nor in an increase of the sample strength when compressed in the [112] direction. However, the interface proved to be a strong obstacle for the non-screw dislocations promoted by the [259] orientation. In this case, dislocation storage – evidenced by large pile-ups – lead to a strong hardening of the specimens. The strengthening contribution of the interface was further affected by the stacking fault energy of the material. While this could be related to the different permeability of the interface, it is more likely the result of a change in cross-slip frequency within the sample volume.

3D Printing Solutions for Microfluidic Chip-To-World Connections.

The connection of microfluidic devices to the outer world by tubes and wires is an underestimated issue. We present methods based on 3D printing to realize microfluidic chip holders with reliable fluidic and electric connections. The chip holders are constructed by microstereolithography, an additive manufacturing technique with sub-millimeter resolution. The fluidic sealing between the chip and holder is achieved by placing O-rings, partly integrated into the 3D-printed structure. The electric connection of bonding pads located on microfluidic chips is realized by spring-probes fitted within the printed holder. Because there is no gluing or wire bonding necessary, it is easy to change the chip in the measurement setup. The spring probes and O-rings are aligned automatically because of their fixed position within the holder. In the case of bioanalysis applications such as cells, a limitation of 3D-printed objects is the leakage of cytotoxic residues from the printing material, cured resin. This was solved by coating the 3D-printed structures with parylene-C. The combination of silicon/glass microfluidic chips fabricated with highly-reliable clean-room technology and 3D-printed chip holders for the chip-to-world connection is a promising solution for applications where biocompatibility, optical transparency and accurate sample handling must be assured. 3D printing technology for such applications will eventually arise, enabling the fabrication of complete microfluidic devices.

High-throughput in situ X-ray screening of and data collection from protein crystals at room temperature and under cryogenic conditions.

Protein crystallography has significantly advanced in recent years, with in situ data collection, in which crystals are placed in the X-ray beam within their growth medium, being a major point of focus. In situ methods eliminate the need to harvest crystals, a previously unavoidable drawback, particularly for often small membrane-protein crystals. Here, we present a protocol for the high-throughput in situ X-ray screening of and data collection from soluble and membrane-protein crystals at room temperature (20-25°C) and under cryogenic conditions. The Mylar in situ method uses Mylar-based film sandwich plates that are inexpensive, easy to make, and compatible with automated imaging, and that show very low background scattering. They support crystallization in microbatch and vapor-diffusion modes, as well as in lipidic cubic phases (LCPs). A set of 3D-printed holders for differently sized patches of Mylar sandwich films makes the method robust and versatile, allows for storage and shipping of crystals, and enables automated mounting at synchrotrons, as well as goniometer-based screening and data collection. The protocol covers preparation of in situ plates and setup of crystallization trials; 3D printing and assembly of holders; opening of plates, isolation of film patches containing crystals, and loading them onto holders; basic screening and data-collection guidelines; and unloading of holders, as well as reuse and recycling of them. In situ plates are prepared and assembled in 1 h; holders are 3D-printed and assembled in ≤90 min; and an in situ plate is opened, and a film patch containing crystals is isolated and loaded onto a holder in 5 min.

Direct Analysis of Aerosolized Chemical Warfare Simulants Captured on a Modified Glass-Based Substrate by "Paper-Spray" Ionization.

Paper spray ionization mass spectrometry offers a rapid alternative platform requiring no sample preparation. Aerosolized chemical warfare agent (CWA) simulants trimethyl phosphate, dimethyl methylphosphonate, and diisopropyl methylphosphonate were captured by passing air through a glass fiber filter disk within a disposable paper spray cartridge. CWA simulants were aerosolized at varying concentrations using an in-house built aerosol chamber. A custom 3D-printed holder was designed and built to facilitate the aerosol capture onto the paper spray cartridges. The air flow through each of the collection devices was maintained equally to ensure the same volume of air sampled across methods. Each approach yielded linear calibration curves with R2 values between 0.98-0.99 for each compound and similar limits of detection in terms of disbursed aerosol concentration. While the glass fiber filter disk has a higher capture efficiency (≈40%), the paper spray method produces analogous results even with a lower capture efficiency (≈1%). Improvements were made to include glass fiber filters as the substrate within the paper spray cartridge consumable. Glass fiber filters were then treated with ammonium sulfate to decrease chemical interaction with the simulants. This allowed for improved direct aerosol capture efficiency (>40%). Ultimately, the limits of detection were reduced to levels comparable to current worker population limits of 1 × 10-6 mg/m3.

Controlled, synchronized actuation of microdroplets by gravity in a superhydrophobic, 3D-printed device

Droplet manipulation over open surfaces allows one to perform assays with a large degree of control and high throughput, making them appealing for applications in drug screening or (bio)analysis. However, the design, manufacturing and operation of these systems comes with high technical requirements. In this study we employ a commercial, low-friction, superhydrophobic coating, Ultra-Ever Dry®, on a 3D-printed microfluidic device. The device features individual droplet compartments, which allow the manipulation of discrete droplets (10–50 μL) actuated by gravity alone. Simply by angling the device to normal in a 3D-printed holder and rocking in a “to and fro”-fashion, a sequence of droplets can be individually transferred to an electrochemical microelectrode detector and then to waste, while preserving the (chronological) order of samples. Multiple biological fluids (i.e. human saliva, urine and rat blood and serum) were successfully tested for compatibility with the device and actuation mechanism, demonstrating low slip angles and high contact angles. Biological matrix (protein) carryover was probed and effectively mitigated by incorporating aqueous rinse droplets as part of the analysis sequence. As a proof-of-concept, the enzyme-coupled, amperometric detection of glucose was carried out on individual rat serum droplets, enabling total analysis in ≈30 min, including calibration. The device is readily customizable, and the integration of droplet generation techniques and other sensor systems for different analytes of interest or applications can be realized in a plug and play fashion.

Microfluidic Platform for the Long-Term On-Chip Cultivation of Mammalian Cells for Lab-On-A-Chip Applications.

Lab-on-a-Chip (LoC) applications for the long-term analysis of mammalian cells are still very rare due to the lack of convenient cell cultivation devices. The difficulties are the integration of suitable supply structures, the need of expensive equipment like an incubator and sophisticated pumps as well as the choice of material. The presented device is made out of hard, but non-cytotoxic materials (silicon and glass) and contains two vertical arranged membranes out of hydrogel. The porous membranes are used to separate the culture chamber from two supply channels for gases and nutrients. The cells are fed continuously by diffusion through the membranes without the need of an incubator and low requirements on the supply of medium to the assembly. The diffusion of oxygen is modelled in order to find the optimal dimensions of the chamber. The chip is connected via 3D-printed holders to the macroscopic world. The holders are coated with Parlyene C to ensure that only biocompatible materials are in contact with the culture medium. The experiments with MDCK-cells show the successful seeding inside the chip, culturing and passaging. Consequently, the presented platform is a step towards Lab-on-a-Chip applications that require long-term cultivation of mammalian cells.

Simultaneous Online Monitoring of Multiple Reactions Using a Miniature Mass Spectrometer.

Advances in chemical sampling using miniature mass spectrometer technology are used to monitor slow reactions at a frequency of ca. 180 h-1 (on the Mini 12) with no sample carryover and with inline derivatization in the case of poorly ionizing compounds. Moreover, we demonstrate high reproducibility with a relative error of less than 10% for major components. Monitoring is enabled using a continuous-flow nanoelectrospray (CF-nESI) probe contained in a custom-built 3D-printed rotary holder. The holder position is automatically set using a stepper motor controlled by a microcontroller. Reaction progress of up to six reactions, including hydrazone formation and Katritzky transamination, can be monitored simultaneously without carryover for several hours.

Fully-Automated μMRI Morphometric Phenotyping of the Tc1 Mouse Model of Down Syndrome.

We describe a fully automated pipeline for the morphometric phenotyping of mouse brains from μMRI data, and show its application to the Tc1 mouse model of Down syndrome, to identify new morphological phenotypes in the brain of this first transchromosomic animal carrying human chromosome 21. We incorporate an accessible approach for simultaneously scanning multiple ex vivo brains, requiring only a 3D-printed brain holder, and novel image processing steps for their separation and orientation. We employ clinically established multi-atlas techniques–superior to single-atlas methods–together with publicly-available atlas databases for automatic skull-stripping and tissue segmentation, providing high-quality, subject-specific tissue maps. We follow these steps with group-wise registration, structural parcellation and both Voxel- and Tensor-Based Morphometry–advantageous for their ability to highlight morphological differences without the laborious delineation of regions of interest. We show the application of freely available open-source software developed for clinical MRI analysis to mouse brain data: NiftySeg for segmentation and NiftyReg for registration, and discuss atlases and parameters suitable for the preclinical paradigm. We used this pipeline to compare 29 Tc1 brains with 26 wild-type littermate controls, imaged ex vivo at 9.4T. We show an unexpected increase in Tc1 total intracranial volume and, controlling for this, local volume and grey matter density reductions in the Tc1 brain compared to the wild-types, most prominently in the cerebellum, in agreement with human DS and previous histological findings.