Name: new application of an atmospheric pressure plasma jet as a neuro-protective agent against glucose deprivation-induced injury of sh-sy5y cells
Uploaded: 2017-10-09T15:00:00
Description: Watch this Scientific Journal Video about New Application of an Atmospheric Pressure Plasma Jet as a Neuro-protective Agent Against Glucose Deprivation-induced Injury of SH-SY5Y Cells at JoVE.com

This work was supported by the Innovation fund of Beijing Neurosurgical Institute (2014-11), National Natural Science Foundation of China (Nos. 11475019 and 81271286) and Beijing Natural Science Foundation (No. 7152027).

1. Preparation of the APPJ device
CAUTION: Please consult all relevant material safety data sheets (MSDS) before use. Please use appropriate safety practices when performing all the&#160;experiments, including the use of a fume hood and personal protective equipment (safety glasses, protective gloves, lab coat, etc.). The protocol requires standard cell handling techniques (sterilizing, cell recovery, cell passaging, cell freezing, cell staining, etc.).
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SH-SY5Y cells are a human neuroblastoma-derived cell line and are widely used as an appropriate cell model for in vitro studies on neurotoxicity or neuroprotection12. The SH-SY5Y cell line was sensitive to glucose deprivation conditions. Cell viability decreased to nearly 50% after 1 h glucose deprivation, which is the optimal cell viability condition for studies of pharmacodynamics. Furthermore, CCK-8 reagent has no cytotoxicity to cells and cells were incubated with CCK-8 reagent for an...

The adult brain almost exclusively uses glucose as a substrate for energy metabolism under normal physiological conditions. The human brain constitutes only 2% of the body weight but consumes approximately 25% of the total glucose within the body1. It is well documented that glucose metabolism dysfunction is one of the major pathological changes during ischemic stroke and various neurodegenerative diseases, including Alzheimer&#39;s disease (AD), Huntington&#39;s disease (HD), and Parkinson&#39;s disease (PD)2,3. The lack of glucose and either impaired glucose uptake or oxidative phosphorylation can directly impact ATP production and further induce neural cell death, which may increase the risk of neuronal dysfunction, suggesting that maintaining cell viability or delaying cell injury after glucose deprivation might be a reasonable approach for treating these diseases. The investigation of neuroprotective effects via glucose modulation, focusing on anti-inflammatory agents, ion channel modulators, free radical scavengers, neurotrophic factors, etc. has been of interest. However, translation of these neuroprotective approaches from bench to clinical practice has not been successful4.
Atmospheric pressure plasma jets (APPJs) are a new kind of atmospheric low temperature gas discharge technology that has attracted the attention of many researchers from multiple disciplines in recent years. APPJs have been used for decades in various biomedical applications such as cancer cell treatment, bacterial inactivation, blood coagulation, wound healing, oral medicine, etc.5,6, due to its emissions of multiple types of reactive nitrogen species (RNS) and reactive oxygen species (ROS) (Figure 1)7. Previous plasma bio-medicine applications mainly focused on the oxidative and/or nitrative stress on bacteria, cells, and tissues8. However, the APPJ could also be a &#34;double-edged sword&#34; since RNS and ROS are important intracellular signaling molecules related to many physiological and pathophysiological processes9. Nitrous oxide (NO) controls a wide range of biological processes and plays a dual role in the human body, especially in central nervous system (CNS). Low levels of NO have shown their neuroprotective activities both in vitro and in vivo via multiple signal pathways10. Our previous study first reported that helium APPJ-induced NO production was involved in the neuroprotective effect of APPJ against oxidative stress-induced injuries11. However, the effects of APPJs on other injuries have not been reported. Therefore, the aim of the present study is to provide an in vitro treatment protocol regarding the neuroprotective applications of helium APPJ on glucose deprivation-induced injury in SH-SY5Y cells. Different from previous studies, our protocol used low-dose plasma treatment for neuroprotective applications without the consequences of excessive plasma-induced injuries, indicating that the APPJ treatment could be potentially used as a novel &#34;NO donor drug&#34; for future research and even for clinical translation. This protocol was also suggested to be used as a cytoprotective application for other cell types with different impairments, but the APPJ treatment conditions should be re-adjusted and the treatment dose must be relatively low.

<table width="911">
	<tbody>
		<tr height="20">
			<td height="20" style="width:216px;height:20px;">SH-SY5Y cell line</td>
			<td style="width:325px;">China Center for Type Culture Collection</td>
			<td style="width:135px;">3111C0001CCC000026</td>
			<td style="width:235px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="width:216px;height:21px;">RPMI 1640 medium</td>
			<td style="width:325px;">Thermo Scientific&#160;</td>
			<td style="width:135px;">21875091</td>
			<td>stored at 4 &#176;C</td>
		</tr>
		<tr height="21">
			<td height="21" style="width:216px;height:21px;">RPMI 1640 medium no glucose</td>
			<td style="width:325px;">Thermo Scientific</td>
			<td style="width:135px;">11879020</td>
			<td>stored at 4 &#176;C</td>
		</tr>
		<tr height="21">
			<td height="21" style="width:216px;height:21px;">fetal calf serum</td>
			<td style="width:325px;">Thermo Scientific</td>
			<td style="width:135px;">16000044</td>
			<td style="width:235px;">stored at -20 &#176;C</td>
		</tr>
		<tr height="21">
			<td height="21" style="width:216px;height:21px;">tripsin-EDTA solution</td>
			<td style="width:325px;">Solarbio</td>
			<td style="width:135px;">T1300</td>
			<td>stored at 4 &#176;C</td>
		</tr>
		<tr height="21">
			<td height="21" style="width:216px;height:21px;">96 wells plate</td>
			<td style="width:325px;">corning</td>
			<td style="width:135px;">3599</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="width:216px;height:21px;">Cell Counting Kit-8 (CCK-8)</td>
			<td style="width:325px;">Dojindo Laboratories</td>
			<td style="width:135px;">CK04</td>
			<td>stored at 4 &#176;C</td>
		</tr>
		<tr height="21">
			<td height="21" style="width:216px;height:21px;">microplate reader</td>
			<td style="width:325px;">Tecan</td>
			<td style="width:135px;">M200 Pro</td>
			<td>for measuring the absorbance at 450 nm</td>
		</tr>
		<tr height="21">
			<td height="21" style="width:216px;height:21px;">High &#8211; voltage Power Amplifier</td>
			<td style="width:325px;">Trek</td>
			<td style="width:135px;">PD06087</td>
			<td>for amplifing the power</td>
		</tr>
		<tr height="21">
			<td height="21" style="width:216px;height:21px;">Function Signal Generator</td>
			<td style="width:325px;">MaZe Electronics Science&#38;Technology</td>
			<td style="width:135px;">AT30120</td>
			<td>for providing the specific signal</td>
		</tr>
		<tr height="21">
			<td height="21" style="width:216px;height:21px;">High &#8211; Voltage Probe</td>
			<td style="width:325px;">Tektronix</td>
			<td style="width:135px;">P6015A</td>
			<td>for detecting high voltage</td>
		</tr>
		<tr height="21">
			<td height="21" style="width:216px;height:21px;">Digital Oscilloscope</td>
			<td style="width:325px;">Tektronix</td>
			<td style="width:135px;">DPO4104B</td>
			<td>for displaying the signal</td>
		</tr>
	</tbody>
</table>

new application of an atmospheric pressure plasma jet as a neuro-protective agent against glucose deprivation-induced injury of sh-sy5y cells

The atmospheric pressure plasma jet (APPJ) has attracted the attention of many researchers from multiple disciplines in recent years because its emissions include multiple types of reactive nitrogen species (RNS) and reactive oxygen species (ROS). Our previous study has shown the cytoprotective effect of the APPJ against oxidative stress-induced injuries. The aim of the present study is to provide a detailed in vitro treatment protocol regarding the neuroprotective applications of helium APPJs on glucose deprivation-induced injury in SH-SY5Y cells. The SH-SY5Y human neuroblastoma-derived cell line was maintained in RPMI 1640 medium supplemented with 15% fetal calf serum. The culture medium was then changed to RPMI 1640 without glucose before APPJ treatment. After a 1 h incubation in a cell incubator, cell viability was determined using Cell Counting Kit 8. The results showed that, compared to the glucose deprivation group, cells treated with APPJ exhibited significantly increased cell viability in a dose-dependent manner, with 8 s/well observed as an optimal dose. Meanwhile, helium flow had no effect on the glucose deprivation-induced cell impairment. Our results indicated that APPJ could be potentially used as a treatment method for the diseases in the central nervous system related to glucose deprivation. This protocol could also be used as a cytoprotective application for other cells with different impairments, but the cell culture and APPJ treatment conditions should be readjusted, and the treatment dose must be relatively low.

Data are expressed as the mean &#177; SD of at least three independent experiments. Group results were analyzed for variance using ANOVA. All analyses were performed using statistical analysis software Prism and p&#60;0.05 was the threshold for statistical significance.
Cell viability was measured after 4 h of CCK-8 incubation. As shown in Figure 3, glucose deprivation reduced the viability of SH-SY5Y cells to 44.1 &#177; 2.6% compared to that of the control group...

Watch this Scientific Journal Video about New Application of an Atmospheric Pressure Plasma Jet as a Neuro-protective Agent Against Glucose Deprivation-induced Injury of SH-SY5Y Cells at JoVE.com

New Application of an Atmospheric Pressure Plasma Jet as a Neuro-protective Agent Against Glucose Deprivation-induced Injury of SH-SY5Y Cells

A protocol for the neuroprotective application of low-dose atmospheric pressure plasma treatment on glucose deprivation-induced SH-SY5Y injuries.

The atmospheric pressure plasma jet (APPJ) has attracted the attention of many researchers from multiple disciplines in recent years because its emissions ...

The overall goal of this treatment method is to provide a detailed in-vitro treatment protocol for neuro-protective applications of helium atmospheric pressure plasma jets on glucose deprivation-induced injury in SH-SY5Y cells. This method can help the key questions in the plasma application field such as plasma medicine. The main advantage of this technique is it acts as a plasma treatment relatively low compared to previous methods for consistent treatments.And we each can prevent excessive plasma-induced injuries. The implications of this technique extend towards therapy of ischemic brain cases because insufficient glucose supply is a common feature of this disease. Though this method can provide insight into new medical applications of plasma it can also be used in other systems such as cardiovascular system.Generally individuals new to this method may struggle because the treatment parameters must be readjusted for different plasma devices. First connect a high voltage power amplifier to a function signal generator that serves as a power supply to provide an AC signal. To record the waveforms of the the applied voltage to a high voltage electrode, connect one end of the high voltage probe to a digital oscilloscope and connect the other end to the power supply.Continually pass helium over the quartz tube of the atmospheric pressure plasma jet device and control the gas flow rate at a stable 1.4 standard liter per minute. Next, turn on the oscilloscope, signal generator and high voltage power amplifier. Rotate the frequency adjustment knob to five kilohertz.Then gradually increase the voltage to a peak to peak value of six kilovolts. After growing an SH-SY5Y human neuroblastoma-derived cell line, carefully aspirate the cell media when the cells reach 85%confluence. Add one milliliter of 0.25%trypsin containing 0.1%EDTA to the cells.After incubating for 15 seconds at room temperature carefully aspirate the trypsin and add two milliliters of RPMI 1640 containing 15%FBS to neutralize. Gently pipette up and down washing the bottom of each well until the SH-SY5Y monolayer is completely detached. Following this, count the cells with a hemocytometer and adjust the cell concentration by adding RPMI 1640 medium containing 15%FBS.Then transfer one hundred microliters of cell suspension to each well of a 96 well plate. Adjust the distance between the nozzle of the quartz tube and the platform where the 96 well plate will be placed to three centimeters to ensure that the beam can touch the culture medium surface. Before treatment, change the culture medium in each well to RPMI 1640 without glucose medium.Now, place the plate under the atmospheric pressure plasma jet nozzle. Then, treat cells in separate wells for zero, one, two, four, eight, and 12 seconds. After treatment, incubate the cells for one hour in a cell incubator at 37 degrees Celsius.Following incubation, add ten microliters of cell counting kit eight solution to each well. Then incubate the cells at 37 degrees Celsius for four hours. Finally measure the absorbance at 450 nanometers with a microplate reader.Glucose deprivation reduced the viability of SH-SY5Y cells to 44.1 plus or minus 2.6 percent compared to that of the control group. The atmospheric pressure plasma jet treatment significantly increased cell viability in a dose-dependent manner at an optimal dose of eight seconds per well. And the cell viability reached 62.27 plus or minus 3.1%Gas flow had no effect on the glucose deprivation-induced cell impairment.Once mastered this technique can be done in 30 hours if it's performed properly. While attempting this procedure it's important to remember to consult all the material safety data sheets before use. After its development, this technology plows a way for researchers in the field of plasma medicine to explore new applications on other kinds of brain injuries and create newer injury models.After watching this video, you should have a good understanding of how to use an atmospheric pressure plasma jet for new practical applications. Don't forget that working with a high voltage power amplifier can be extremely hazardous and precautions such as not touching the high voltage lines should always be taken when performing this procedure.

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Engineering

Synthesis and Microdiffraction at Extreme Pressures and Temperatures

High pressure compounds and polymorphs are investigated for a broad range of purposes such as determine structures and processes of deep planetary interiors, design materials with novel properties, understand the mechanical behavior of materials exposed to very high stresses as in explosions or impacts. Synthesis and structural analysis of materials at extreme conditions of pressure and temperature entails remarkable technical challenges. In the laser heated diamond anvil cell (LH-DAC), very high pressure is generated between the tips of two opposing diamond anvils forced against each other; focused infrared laser beams, shined through the diamonds, allow to reach very high temperatures on samples absorbing the laser radiation. When the LH-DAC is installed in a synchrotron beamline that provides extremely brilliant x-ray radiation, the structure of materials under extreme conditions can be probed in situ. LH-DAC samples, although very small, can show highly variable grain size, phase and chemical composition. In order to obtain the high resolution structural analysis and the most comprehensive characterization of a sample, we collect diffraction data in 2D grids and combine powder, single crystal and multigrain diffraction techniques. Representative results obtained in the synthesis of a new iron oxide, Fe4O5 1 will be shown.

The laser heated diamond anvil cell combined with synchrotron micro-diffraction techniques allows researchers to explore the nature and properties of new phases of matter at extreme pressure and temperature (PT) conditions. Heterogeneous samples can be characterized in situ under high pressure by 2D mapping and combined powder, single-crystal and multigrain diffraction approaches.

High pressure compounds and polymorphs are investigated for a broad range of purposes such as determine structures and processes of deep planetary ...

Visualization of High Speed Liquid Jet Impaction on a Moving Surface

Two apparatuses for examining liquid jet impingement on a high-speed moving surface are described: an air cannon device (for examining surface speeds between 0 and 25 m/sec) and a spinning disk device (for examining surface speeds between 15 and 100 m/sec). The air cannon linear traverse is a pneumatic energy-powered system that is designed to accelerate a metal rail surface mounted on top of a wooden projectile. A pressurized cylinder fitted with a solenoid valve rapidly releases pressurized air into the barrel, forcing the projectile down the cannon barrel. The projectile travels beneath a spray nozzle, which impinges a liquid jet onto its metal upper surface, and the projectile then hits a stopping mechanism. A camera records the jet impingement, and a pressure transducer records the spray nozzle backpressure. The spinning disk set-up consists of a steel disk that reaches speeds of 500 to 3,000 rpm via a variable frequency drive (VFD) motor. A spray system similar to that of the air cannon generates a liquid jet that impinges onto the spinning disc, and cameras placed at several optical access points record the jet impingement. Video recordings of jet impingement processes are recorded and examined to determine whether the outcome of impingement is splash, splatter, or deposition. The apparatuses are the first that involve the high speed impingement of low-Reynolds-number liquid jets on high speed moving surfaces. In addition to its rail industry applications, the described technique may be used for technical and industrial purposes such as steelmaking and may be relevant to high-speed 3D printing.

Two experimental devices for examining liquid jet impingement on a high-speed moving surface are described: an air cannon device and a spinning disk device. The apparatuses are used to determine optimal approaches to the application of liquid friction modifier (LFM) onto rail tracks for top-of-rail friction control.

Two apparatuses for examining liquid jet impingement on a high-speed moving surface are described: an air cannon device (for examining surface speeds ...

How to Ignite an Atmospheric Pressure Microwave Plasma Torch without Any Additional Igniters

This movie shows how an atmospheric pressure plasma torch can be ignited by microwave power with no additional igniters. After ignition of the plasma, a stable and continuous operation of the plasma is possible and the plasma torch can be used for many different applications. On one hand, the hot (3,600 K gas temperature) plasma can be used for chemical processes and on the other hand the cold afterglow (temperatures down to almost RT) can be applied for surface processes. For example chemical syntheses are interesting volume processes. Here the microwave plasma torch can be used for the decomposition of waste gases which are harmful and contribute to the global warming but are needed as etching gases in growing industry sectors like the semiconductor branch. Another application is the dissociation of CO2. Surplus electrical energy from renewable energy sources can be used to dissociate CO2 to CO and O2. The CO can be further processed to gaseous or liquid higher hydrocarbons thereby providing chemical storage of the energy, synthetic fuels or platform chemicals for the chemical industry. Applications of the afterglow of the plasma torch are the treatment of surfaces to increase the adhesion of lacquer, glue or paint, and the sterilization or decontamination of different kind of surfaces. The movie will explain how to ignite the plasma solely by microwave power without any additional igniters, e.g., electric sparks. The microwave plasma torch is based on a combination of two resonators &#8212; a coaxial one which provides the ignition of the plasma and a cylindrical one which guarantees a continuous and stable operation of the plasma after ignition. The plasma can be operated in a long microwave transparent tube for volume processes or shaped by orifices for surface treatment purposes.

This movie shows how an atmospheric plasma torch can be ignited by microwaves with no additional igniters and provides a stable and continuous plasma operation suitable for plenty of applications.

This movie shows how an atmospheric pressure plasma torch can be ignited by microwave power with no additional igniters. After ignition of the plasma, a ...

Three-dimensional Particle Tracking Velocimetry for Turbulence Applications: Case of a Jet Flow

3D-PTV is a quantitative flow measurement technique that aims to track the Lagrangian paths of a set of particles in three dimensions using stereoscopic recording of image sequences. The basic components, features, constraints and optimization tips of a 3D-PTV topology consisting of a high-speed camera with a four-view splitter are described and discussed in this article. The technique is applied to the intermediate flow field (5 &#60;x/d &#60;25) of a circular jet at Re &#8776; 7,000. Lagrangian flow features and turbulence quantities in an Eulerian frame are estimated around ten diameters downstream of the jet origin and at various radial distances from the jet core. Lagrangian properties include trajectory, velocity and acceleration of selected particles as well as curvature of the flow path, which are obtained from the Frenet-Serret equation. Estimation of the 3D velocity and turbulence fields around the jet core axis at a cross-plane located at ten diameters downstream of the jet is compared with literature, and the power spectrum of the large-scale streamwise velocity motions is obtained at various radial distances from the jet core.

A three-dimensional particle tracking velocimetry (3D-PTV) system based on a high-speed camera with a four-view splitter is described here. The technique is applied to a jet flow from a circular pipe in the vicinity of ten diameters downstream at Reynolds number Re &#8776; 7,000.

3D-PTV is a quantitative flow measurement technique that aims to track the Lagrangian paths of a set of particles in three dimensions using stereoscopic ...

Applying X-ray Imaging Crystal Spectroscopy for Use as a High Temperature Plasma Diagnostic

X-ray spectra provide a wealth of information on high temperature plasmas; for example electron temperature and density can be inferred from line intensity ratios. By using a Johann spectrometer viewing the plasma, it is possible to construct profiles of plasma parameters such as density, temperature, and velocity with good spatial and time resolution. However, benchmarking atomic code modeling of X-ray spectra obtained from well-diagnosed laboratory plasmas is important to justify use of such spectra to determine plasma parameters when other independent diagnostics are not available. This manuscript presents the operation of the High Resolution X-ray Crystal Imaging Spectrometer with Spatial Resolution (HIREXSR), a high wavelength resolution spatially imaging X-ray spectrometer used to view hydrogen- and helium-like ions of medium atomic number elements in a tokamak plasma. In addition, this manuscript covers a laser blow-off system that can introduce such ions to the plasma with precise timing to allow for perturbative studies of transport in the plasma.

X-ray spectra provide a wealth of information on high temperature plasmas. This manuscript presents the operation of a high wavelength resolution spatially imaging X-ray spectrometer used to view hydrogen- and helium-like ions of medium atomic number elements in a tokamak plasma.

X-ray spectra provide a wealth of information on high temperature plasmas; for example electron temperature and density can be inferred from line ...

An Atmospheric Pressure Plasma Setup to Investigate the Reactive Species Formation

Non-thermal atmospheric pressure (&#39;cold&#39;) plasmas have received increased attention in recent years due to their significant biomedical potential. The reactions of cold plasma with the surrounding atmosphere yield a variety of reactive species, which can define its effectiveness. While efficient development of cold plasma therapy requires kinetic models, model benchmarking needs empirical data. Experimental studies of the source of reactive species detected in aqueous solutions exposed to plasma are still scarce. Biomedical plasma is often operated with He or Ar feed gas, and a specific interest lies in investigation of the reactive species generated by plasma with various gas admixtures (O2, N2, air, H2O vapor, etc.) Such investigations are very complex due to difficulties in controlling the ambient atmosphere in contact with the plasma effluent. In this work, we addressed common issues of &#39;high&#39; voltage kHz frequency driven plasma jet experimental studies. A reactor was developed allowing the exclusion of ambient atmosphere from the plasma-liquid system. The system thus comprised the feed gas with admixtures and the components of the liquid sample. This controlled atmosphere allowed the investigation of the source of the reactive oxygen species induced in aqueous solutions by He-water vapor plasma. The use of isotopically labelled water allowed distinguishing between the species originating in the gas phase and those formed in the liquid. The plasma equipment was contained inside a Faraday cage to eliminate possible influence of any external field. The setup is versatile and can aid in further understanding the cold plasma-liquid interactions chemistry.

An experimental setup was created for the helium-operated kHz frequency plasma jet. The setup includes a cage for the plasma power supply and jet and an in-house built reactor to monitor plasma-induced reactive species without the interference of the ambient atmosphere.

Non-thermal atmospheric pressure (&#39;cold&#39;) plasmas have received increased attention in recent years due to their significant biomedical potential. ...

Atmospheric Pressure Fabrication of Large-Sized Single-Layer Rectangular SnSe Flakes

Tin selenide (SnSe) belongs to the family of layered metal chalcogenide materials with a buckled structure like phosphorene, and has shown potential for applications in two-dimensional nanoelectronics devices. Although many methods to synthesize SnSe nanocrystals have been developed, a simple way to fabricate large-sized single-layer SnSe flakes remains a great challenge. Herein, we show the experimental method to directly grow large-sized single-layer rectangular SnSe flakes on commonly used SiO2/Si insulating substrates using a straightforward two-step fabrication method in an atmospheric pressure quartz tube furnace system. The single-layer rectangular SnSe flakes with an average thickness of ~6.8 &#197; and lateral dimensions of about 30 &#181;m &#215; 50 &#181;m were fabricated by a combination of vapor transport deposition technique and nitrogen etching route. We characterized the morphology, microstructure, and electrical properties of the rectangular SnSe flakes and obtained excellent crystallinity and good electronic properties. This article about the two-step fabrication method can help researchers grow other similar two-dimensional, large-sized, single-layer materials using an atmospheric pressure system.

A protocol is presented demonstrating a two-step fabrication technique to grow large-sized single-layer rectangular shaped SnSe flakes on low-cost SiO2/Si dielectrics wafers in an atmospheric pressure quartz tube furnace system.

Tin selenide (SnSe) belongs to the family of layered metal chalcogenide materials with a buckled structure like phosphorene, and has shown potential for ...

Magnet Assisted Composite Manufacturing: A Flexible New Technique for Achieving High Consolidation Pressure in Vacuum Bag/Lay-Up Processes

This work demonstrates a protocol to improve the quality of composite laminates fabricated by wet lay-up vacuum bag processes using the recently developed magnet assisted composite manufacturing (MACM) technique. In this technique, permanent magnets are utilized to apply a sufficiently high consolidation pressure during the curing stage. To enhance the intensity of the magnetic field, and thus, to increase the magnetic compaction pressure, the magnets are placed on a magnetic top plate. First, the entire procedure of preparing the composite lay-up on a magnetic bottom steel plate using the conventional wet lay-up vacuum bag process is described. Second, placement of a set of Neodymium-Iron-Boron permanent magnets, arranged in alternating polarity, on the vacuum bag is illustrated. Next, the experimental procedures to measure the magnetic compaction pressure and volume fractions of the composite constituents are presented. Finally, methods used to characterize microstructure and mechanical properties of composite laminates are discussed in detail. The results prove the effectiveness of the MACM method in improving the quality of wet lay-up vacuum bag laminates. This method does not require large capital investment for tooling or equipment and can also be used to consolidate geometrically complex composite parts by placing the magnets on a matching top mold positioned on the vacuum bag.

A new technique for applying consolidation pressure on the vacuum bag lay-up to fabricate composite laminates is described. The goal of this protocol is to develop a simple, cost-effective technique to improve the quality of laminates fabricated by the wet lay-up vacuum bag method.

This work demonstrates a protocol to improve the quality of composite laminates fabricated by wet lay-up vacuum bag processes using the recently developed ...

Nerve Stem Cell Differentiation by a One-step Cold Atmospheric Plasma Treatment In Vitro

As the development of physical plasma technology, cold atmospheric plasmas (CAPs) have been widely investigated in decontamination, cancer treatment, wound healing, root canal treatment, etc., forming a new research field named plasma medicine. Being a mixture of electrical, chemical, and biological reactive species, CAPs have shown their abilities to enhance nerve stem cells differentiation both in vitro and in vivo and are becoming a promising way for neurological disease treatment in the future. The much more exciting news is that using CAPs may realize one-step, and safely directed, differentiation of neural stem cells (NSCs) for tissue transportation. We demonstrate here the detailed experimental protocol of using a self-made CAP jet device to enhance NSC differentiation in C17.2-NSCs and primary rat neural stem cells, as well as observing the cell fate by inverted and fluorescence microscopy. With the help of immunofluorescence staining technology, we found both the NSCs showed an accelerated differential rate than the untreated group, and ~75% of the NSCs selectively differentiated into neurons, which are mainly mature, cholinergic, and motor neurons.

Nerve Stem Cell Differentiation by a One-step Cold Atmospheric Plasma Treatment In Vitro

This protocol aims to provide detailed experimental steps of a cold atmospheric plasma treatment on neural stem cells and immunofluorescence detection for differentiation enhancement.

As the development of physical plasma technology, cold atmospheric plasmas (CAPs) have been widely investigated in decontamination, cancer treatment, ...

Recording Ultra-Realistic Full-Color Analog Holograms for Use in a Moving Hologram Display

This paper demonstrates a method to record a set of twelve ultra-realistic full-color analog holograms presenting the same brightness, transparency and homogeneous colors for the fabrication of a Fantatrope, a dynamic holographic 3D display, without the need for special viewing aids. The method involves the use of 3D printer technology, a single-beam full-color Denisyuk optical setup with three low-power lasers (red, green, and blue) and an iso-panchromatic high-sensitive silver-halide holographic emulsion specially designed for recording analog holograms without any diffusion. A cyclic animation is created with a 3D computer graphics program and different elements are 3D printed to form models for the holograms. Holograms are recorded with a full-color holographic setup and developed using two simple chemical baths. To prevent any emulsion thickness variations, the holograms are sealed with optical glue. Results confirm that all holograms recorded with this protocol present the same characteristics, which allow them to be used in the Fantatrope.

We present a protocol to record a set of ultra-realistic full-color analog holograms, showing the same brightness, transparency, and homogeneous colors, on ultra-fine-grain silver-halide holographic emulsions for the fabrication of a dynamic holographic 3D display.

This paper demonstrates a method to record a set of twelve ultra-realistic full-color analog holograms presenting the same brightness, transparency and ...