How to Control the Microfluidic Flow and Separate the Magnetic and Non-Magnetic Particles in the Runner of a Disc

Biochips play an important role in both medical and food industry safety testing. Moreover, magnetic activated cell sorting is a well-established technology for biochip development. However, biochips need to be manufactured by precision instruments, resulting in the high cost of biochips. Therefore, this study used magnetic-activation and mechanics theories to create a novel disc that could manipulate the microfluidic flow, mixing, reaction, and separation on the runner of the disc. The goal of the research was to apply in the field of biomedical detection systems to reduce the cost of biochips and simplify the operation process. The simulation and experimental investigation showed that the pattern of the reaction chamber was stomach-shaped and the reservoir chamber was rectangular-shaped on the disc. The microfluid could be controlled to flow to the reaction chamber from the buffer and sample chamber when the disc spun at 175~200 rpm within three minutes. This was defined as the first setting mode. The microfluid could then be controlled to flow to the reservoir chamber from the reaction chamber when the disc spun at 225 rpm within five to ten minutes. This was defined as the second setting mode. This verified that the pattern design of the disc was optimized for control of the microfluid flow, mixing, reaction, and separation in the runner of the disc by different setting modes.

HIDUNG ELEKTRONIK BERBASIS SENSOR GAS MOS UNTUK KARAKTERISASI KEMATANGAN BUAH KELAPA SAWIT

The ripeness of oil palm fruits is one of the key factors for crude palm oil qualities. Recently, electronic nose systems have been developed intensively for fruit quality assessment which relates odors to ripeness levels. This study developed an electronic nose system to characterize the ripeness levels of oil palm fruits using output voltage of each sensor and fruit hardness. The system consisted of a sensor chamber and a sample chamber. The sensor chamber consisted of eight MOS gas sensor modules of MQ series. Samples were oil palm fruits taken from oil palm fresh fruit bunches (FFB) which were previously categorized traditionally into unripe, ripe, over ripe, peeled and put into the sample chamber. Some of the fruits were also used for hardness measurement. To quantify the output voltages for each sensor, integrated trapezoid areas were calculated and related to the fruit hardness values. The results showed a significant voltage difference of each sensor for the three ripeness levels. Only four out of eight sensors showed significantly higher voltages. Three sensors which can significantly differentiate the ripeness levels are MQ3, MQ5, and MQ135 which MQ135 is the best. This shows that the electronic nose is potential for oil palm fruits. Keywords: electronic nose, fruit hardness, MOS gas sensor, oil palm fruit, ripeness

Novel experimental setup for megahertz X-ray diffraction in a diamond anvil cell at the High Energy Density (HED) instrument of the European X-ray Free-Electron Laser (EuXFEL)

The high-precision X-ray diffraction setup for work with diamond anvil cells (DACs) in interaction chamber 2 (IC2) of the High Energy Density instrument of the European X-ray Free-Electron Laser is described. This includes beamline optics, sample positioning and detector systems located in the multipurpose vacuum chamber. Concepts for pump–probe X-ray diffraction experiments in the DAC are described and their implementation demonstrated during the First User Community Assisted Commissioning experiment. X-ray heating and diffraction of Bi under pressure, obtained using 20 fs X-ray pulses at 17.8 keV and 2.2 MHz repetition, is illustrated through splitting of diffraction peaks, and interpreted employing finite element modeling of the sample chamber in the DAC.

Disease Modeling with 3D Cell-Based Assays Using a Novel Flowchip System and High-Content Imaging

There is an increasing interest in using three-dimensional (3D) cell structures for modeling tumors, organs, and tissue to accelerate translational research. We describe here a novel automated organoid assay system (the Pu·MA System) combined with microfluidic-based flowchips that can facilitate 3D cell-based assays. The flowchip is composed of sample wells, which contain organoids, connected to additional multiple wells that can hold various assay reagents. Organoids are positioned in a protected chamber in sample wells, and fluids are exchanged from side reservoirs using pressure-driven flow. Media exchange, sample staining, wash steps, and other processes can be performed without disruption to or loss of 3D sample. The bottom of the sample chamber is thin, optically clear plastic compatible with high-content imaging (HCI). The whole system can be kept in an incubator, allowing long-term cellular assays to be performed. We present two examples of use of the system for biological research. In the first example, cytotoxicity effects of anticancer drugs were evaluated on HeLa and HepG2 spheroids using HCI and vascular endothelial growth factor expression. In the second application, the flowchip system was used for the functional evaluation of Ca2+ oscillations in neurospheroids. Neurospheres were incubated with neuroactive compounds, and neuronal activity was assessed using Ca2+-sensitive dyes and fast kinetic fluorescence imaging. This novel assay system using microfluidics enables automation of 3D cell-based cultures that mimic in vivo conditions, performs multidosing protocols and multiple media exchanges, provides gentle handling of spheroids and organoids, and allows a wide range of assay detection modalities.

A Dose-Response Model and D10-Value for Mycobacterium tuberculosis Exposed to Dosimetrically Verified Ionizing Radiation

Techniques for pathogen inactivation have been employed by laboratories to help ease the financial, physical, and health strains associated with (A)BSL-3 work. Exposure to radiation is the most common and useful of these methods to inactivate pathogens grown in large-scale culture. While robust protocols exist for radiation exposure techniques, there are variances in methods used to determine the radiation dose and dose rate required to inactivate pathogens. Furthermore, previous studies often do not include radiation dosimetry verification or address corresponding dosimetry uncertainties for dose response-assays. Accordingly, this study was conducted with the purpose of completing a dosimetry assessment of the radiation field within the sample chamber of a sealed source irradiator, to subsequently determine the radiation dose required to inactivate pathogenic cultures. Physical dosimetry techniques (Fricke dosimetry, ion chamber measurements, and measurements with thermoluminescent dosimeters) were used to measure dose rate and rate variances within the sample chamber. By comparing the variances between the dosimetry methodologies and measurements, an estimated dose rate within the sample chamber was determined. The results of the dosimetry evaluation were used to determine the radiation dose samples of Mycobacterium tuberculosis received, to accurately associate biological markers of inactivation to specific doses of ionizing radiation. A D10 value and dose-response curve were developed to describe the inactivation of Mtb from increasing doses of ionizing radiation. The D10 value is experimentally relevant for comparative analysis and potentially provides a biological baseline for inactivation verification. This methodology can also easily be translated to other pathogen models.ImportanceThis work set out to give us a better understanding of how much radiation is required to inactivate Mycobacterium tuberculosis, the bacteria that causes tuberculosis disease. Radiation dose from a source is not something that can just be inputted, it must be calculated, so we also determined the approximate dose from the source to address ambiguities that had previously existed while inactivating microbes. We were able to generate an accurate description of inactivation of Mycobacterium tuberculosis by correlating it with a value representing 90% death of the treated cells. We also unexpectedly discovered that very low levels of radiation increase certain activity within the cell. This is important because it allows us to better understand how radiation kills Mycobacterium tuberculosis, and gives us a value to compare to other organisms. It also offers other researchers a method to use under their own specific conditions.

Sample chamber for synchrotron based in-situ X-ray diffraction experiments under electric fields and temperatures between 100 K and 1250 K

Many scientific questions require X-ray experiments conducted at varying temperatures, sometimes combined with the application of electric fields. Here, a customized sample chamber developed for beamlines P23 and P24 of PETRA III at DESY to suit these demands is presented. The chamber body consists mainly of standard vacuum parts housing the heater/cooler assembly supplying a temperature range of 100 K to 1250 K and an xyz manipulator holding an electric contact needle for electric measurements at both high voltage and low current. The chamber is closed by an exchangeable hemispherical dome offering all degrees of freedom for single-crystal experiments within one hemisphere of solid angle. The currently available dome materials (PC, PS, PEEK polymers) differ in their absorption and scattering characteristics, with PEEK providing the best overall performance. The article further describes heating and cooling capabilities, electric characteristics, and plans for future upgrades of the chamber. Examples of applications are discussed.

Phase Stability and Vibrational Properties of Iron-Bearing Carbonates at High Pressure

The spin transition of iron can greatly affect the stability and various physical properties of iron-bearing carbonates at high pressure. Here, we reported laser Raman measurements on iron-bearing dolomite and siderite at high pressure and room temperature. Raman modes of siderite FeCO3 were investigated up to 75 GPa in the helium (He) pressure medium and up to 82 GPa in the NaCl pressure medium, respectively. We found that the electronic spin-paring transition of iron in siderite occurred sharply at 42–44 GPa, consistent with that in the neon (Ne) pressure medium in our previous study. This indicated that the improved hydrostaticity from Ne to He had minimal effects on the spin transition pressure. Remarkably, the spin crossover of siderite was broadened to 38–48 GPa in the NaCl pressure medium, due to the large deviatoric stress in the sample chamber. In addition, Raman modes of iron-bearing dolomite Ca1.02Mg0.76Fe0.20Mn0.02(CO3)2 were explored up to 58 GPa by using argon as a pressure medium. The sample underwent phase transitions from dolomite-Ⅰ to -Ⅰb phase at ~8 GPa, and then to -Ⅱ at ~15 and -Ⅲb phase at 36 GPa, while no spin transition was observed in iron-bearing dolomite up to 58 GPa. The incorporation of FeCO3 by 20 mol% appeared to marginally decrease the onset pressures of the three phase transitions aforementioned for pure dolomite. At 55–58 GPa, the ν1 mode shifted to a lower frequency at ~1186 cm−1, which was likely associated with the 3 + 1 coordination in dolomite-Ⅲb. These results shed new insights into the nature of iron-bearing carbonates at high pressure.

Microfluidic flow-cell with passive flow control for microscopy applications

We present a fast, inexpensive and robust technique for constructing thin, optically transparent flow-cells with pump-free flow control. Using layers of glass, patterned adhesive tape and polydimethylsiloxane (PDMS) connections, we demonstrate the fabrication of planar devices with chamber height as low as 25 μm and with millimetre-scale (x,y) dimensions for wide-field microscope observation. The method relies on simple benchtop equipment and does not require microfabrication facilities, glass drilling or other workshop infrastructure. We also describe a gravity perfusion system that exploits the strong capillary action in the flow chamber as a passive limit-valve. Our approach allows simple sequential sample exchange with controlled flow rates, sub-5 μL sample chamber size and zero dead volume. We demonstrate the system in a single-molecule force spectroscopy experiment using magnetic tweezers.

Diamond B23 CD Imaging of Thin Films of Chiral Materials or Achiral Polymers Coated with Chiral Molecules

The novel vertical sample chamber, developed at the B23 beamline for synchrotron radiation circular dichroism (SRCD), has enabled the Diamond User community to conduct different types of experiments from high throughput CD of protein and DNA folding using 96-well multiplates to CD imaging at high spatial resolution. Here, we present the application of CD imaging to large areas of achiral polymer PVA films doped with D-dopa to assess the chiral homogeneity of the film preparation with potential antimicrobial property. Synopsis: CDi application of Diamond B23 SRCD beamline.

An ultra-low magnetic field thermal demagnetizer for high-precision paleomagnetism

Thermal demagnetization furnaces are widely used paleomagnetic facilities for progressive removal of naturally acquired magnetic remanence or the imparting of well-controlled laboratory magnetization. An ideal thermal demagnetizer should maintain “zero” magnetic field in the sample chamber during thermal treatments. However, magnetic field noises, including the residual magnetic fields of the construction material and the induced fields caused by the alternating current (AC) in the heating element are always present, which can contaminate the paleomagnetic results at the elevated temperatures or especially for the magnetically weak samples. Here, we designed a new structure of heating wire named “straight core solenoid” to develop a new demagnetization furnace with ultra-low magnetic field noise. Simulation and practical measurements show that the heating current magnetic field can be greatly reduced by using the new technology. Thermal demagnetization experiments demonstrate that the new demagnetizer can yield low noise results even for weakly magnetic samples.