The authors gratefully acknowledge the Department of Chemistry and the Graduate College at Missouri State University for their support of this project. We also acknowledge the Electron Microscopy Laboratory at the Frederick National Laboratory for Cancer Research for use of their transmission electron microscope and carbon-coated grids.

1. Synthesis of Indium Phosphide/Zinc Sulfide (InP/ZnS) Quantum Dots
<ol>
	<li>Synthesis of Indium Phosphide (InP) Quantum Dot Cores
	<ol>
		<li>Fit a 100 ml round bottom, 3-neck, flask with a 12-inch condenser. Add 30 ml oleylamine (OLA), 0.398 g indium (III) chloride (InCl3), 0.245 g zinc (II) chloride (ZnCl2) and stir while evacuating at RT using a vacuum for 1 hr. The solution should appear colorless with a white precipitate.</li>
		<li>Using a heating mantle with a thermocoup...

<ol>
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This protocol details the synthesis of highly fluorescent InP/ZnS QDs that can be used in many biological systems. The QD products synthesized here exhibited a single fluorescence emission peak centered at 600 nm with a FWHM of 73 nm (Figure 1), which is comparable to other previously described syntheses12. Reaction time and reaction temperature are extremely crucial steps due to their profound effect on QD synthesis quality and repeatability. After solubilization in water, the QDs were determ...

A correction was made to: <a href="http://www.jove.com/video/53684/synthesis-cd-free-inpzns-quantum-dots-suitable-for-biomedical" target="_blank">Synthesis of Cd-free InP/ZnS Quantum Dots Suitable for Biomedical Applications</a>. There was an error with an author's given name. The author's name was corrected to:
Katye M. Fichter
from:
Kathryn M. Fichter

A correction was made to: <a href="http://www.jove.com/video/53684/synthesis-cd-free-inpzns-quantum-dots-suitable-for-biomedical" target="_blank">Synthesis of Cd-free InP/ZnS Quantum Dots Suitable for Biomedical Applications</a>.

Erratum: Synthesis of Cd-free InP/ZnS Quantum Dots Suitable for Biomedical Applications

Quantum dots (QDs) are semiconducting nanocrystals that exhibit fluorescent properties when irradiated with light1. Due to their small size (2-5 nm), which is similar to many larger biomolecules, and ease of biofunctionalization, QDs are an extremely attractive tool for biomedical applications. They have found use in biological labeling, single-molecule live-cell imaging, drug delivery, in vivo imaging, pathogen detection, and cell tracking, among many other uses2-8.
Cd-based QDs have been most commonly used in biomedical applications because of their intense fluorescence and narrow emission peak widths9. However, concerns have been raised due to potential toxicity of Cd2+ ions10 that may be released through degradation of the nanoparticle. Recently, InP-based QDs have been explored as an alternative to Cd-based QDs because they maintain many fluorescence characteristics of Cd-based QDs and may be more biocompatible11. Cd-based QDs have been found to be significantly more toxic than InP-based QDs in in vitro assays at concentrations as low as 10 pM, after only 48 hr11.
The fluorescence emission color of QDs is size-tunable1. That is, as the size of the QD increases, the fluorescence emission is red-shifted. The size and size dispersity of the QD products can be modified by changing the temperature, reaction duration, or precursor concentration conditions during the reaction12. While the emission peak of InP QDs is typically broader and less intense than Cd-based QDs, InP QDs can be made in a large variety of colors designed to avoid spectral overlap, and are sufficiently intense for most biomedical applications12. The synthesis detailed in this protocol yields QDs with a red emission peak centered at 600 nm.
Several steps are taken after synthesis of the QD cores to maintain the optical integrity of the QDs and to make them compatible for biological applications. The surface of the QD core must be protected from oxidation or surface defects that may cause quenching; therefore, a ZnS shell is coated over the core to produce InP/ZnS (core/shell) QDs13. This coating has been shown to protect the photoluminescence of the QD product. The presence of zinc ions during InP QD synthesis has been shown to limit surface defects, as well as decrease size distribution12. Even with the presence of Zn2+ in the reaction medium, synthesis of InZnP are highly unlikely12. After coating, resulting InP/ZnS QDs are coated in hydrophobic ligands such as trioctylphosphine oxide (TOPO) or oleylamine12,14. An amphiphilic polymer can interact with hydrophobic ligands on the QD surface as well as bulk water molecules to impart water solubility15. Amphiphilic polymers with carboxylate chemical groups can be used as &#34;chemical handles&#34; to further functionalize the QDs.
This protocol details the synthesis and functionalization of water-soluble InP/ZnS QDs with very intense fluorescence emission and relatively small size-dispersity. These QDs are potentially less toxic than commonly used CdSe/ZnS QDs. Herein, the synthesis of InP/ZnS QDs provides a practical alternative to Cd-based QDs for biomedical applications.

<table border="0" cellpadding="0" cellspacing="0" width="725">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:232px;">Oleylamine</td>
			<td style="width:92px;">Acros</td>
			<td style="width:119px;">129540010</td>
			<td style="width:283px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:232px;">Zinc (II) chloride</td>
			<td style="width:92px;">Sigma</td>
			<td style="width:119px;">030-003-00-2</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:232px;">Indium (III) chloride</td>
			<td style="width:92px;">Chem-Impex&#160;</td>
			<td style="width:119px;">24560</td>
			<td></td>
		</tr>
		<tr height="27">
			<td height="27" style="height:27px;width:232px;">Tris(dimethylamino)phosphine</td>
			<td style="width:92px;">Encompass</td>
			<td style="width:119px;">50-901-10500</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:232px;">1-dodecanethiol</td>
			<td style="width:92px;">Acros</td>
			<td style="width:119px;">117625000</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:232px;">Hexanes</td>
			<td style="width:92px;">Fisher Sci</td>
			<td style="width:119px;">H292-4</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:232px;">Acetone</td>
			<td style="width:92px;">TransChemical</td>
			<td style="width:119px;">UN 1090</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:232px;">Zinc Stearate</td>
			<td style="width:92px;">Aldrich Chem</td>
			<td style="width:119px;">307564-1KG</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:232px;">Tetrahydrofuran</td>
			<td style="width:92px;">Acros</td>
			<td style="width:119px;">34845-0010</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:232px;">Molecular Water</td>
			<td style="width:92px;">Fisher Sci</td>
			<td style="width:119px;">BP2470-1</td>
			<td></td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:232px;">Poly(maleic anhyrdride-alt-1-tetradecene), 3-(dimethylamino)-1-propylamine derivative</td>
			<td style="width:92px;">Sigma</td>
			<td style="width:119px;">90771-1G</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:232px;">Boric acid</td>
			<td style="width:92px;">Fisher Sci</td>
			<td style="width:119px;">BP168-500</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:232px;">Sodium Tetraborate Decahydrate</td>
			<td style="width:92px;">Fisher Sci</td>
			<td style="width:119px;">BP175-500</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:232px;">Rhodamine B</td>
			<td style="width:92px;">Aldrich Chem</td>
			<td style="width:119px;">R95-3</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:232px;">Nitrogen gas</td>
			<td style="width:92px;">Airgas</td>
			<td style="width:119px;">UN1066</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:232px;">Trypan blue</td>
			<td style="width:92px;">Thermo Sci</td>
			<td style="width:119px;">SV30084.01</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:232px;">3 mL plastic Luer-lock syringe</td>
			<td style="width:92px;">BD</td>
			<td style="width:119px;">309657</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:232px;">Luer-lock Needle</td>
			<td style="width:92px;">Air-Tite</td>
			<td style="width:119px;">8300014471</td>
			<td>4 inch, 22 gauge</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:232px;">50 mL polypropyene centrifuge tube</td>
			<td style="width:92px;">Falcon</td>
			<td style="width:119px;">352098</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:232px;">250 mL centrifuge bottle</td>
			<td style="width:92px;">Thermo Sci</td>
			<td>05-562-23</td>
			<td>Nalgene PPCO</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:232px;">5 mL centrifuge tubes</td>
			<td style="width:92px;">Argos-Tech</td>
			<td style="width:119px;">T2076</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:232px;">1.5 mL microcentrifuge tubes</td>
			<td style="width:92px;">Bio Plas</td>
			<td style="width:119px;">4150</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:232px;">0.1 &#956;m Syringe filter</td>
			<td style="width:92px;">Whatman</td>
			<td style="width:119px;">6786-1301</td>
			<td>Puradisc 13 mm nylon filter</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:232px;">Slide-A-Lyzer MINI Dialysis Unit</td>
			<td style="width:92px;">Thermo Sci</td>
			<td style="width:119px;">69590</td>
			<td>20,000 MWCO</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:232px;">Rotary Evaporator</td>
			<td style="width:92px;">Heidolph</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:232px;">Centrifuge 5072</td>
			<td style="width:92px;">Eppendorf</td>
			<td style="width:119px;"></td>
			<td>Swinging Bucket with 50 mL tube adapters</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:232px;">Lambda 650 UV/VIS Spectrometer</td>
			<td style="width:92px;">Perkin Elmer</td>
			<td style="width:119px;"></td>
			<td style="width:283px;">UV-Vis Spectrophotometer</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:232px;">LS 55 Fluorescence Spectrometer</td>
			<td style="width:92px;">Perkin Elmer</td>
			<td style="width:119px;"></td>
			<td style="width:283px;">Fluorometer</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:232px;">Axio Observer.A1</td>
			<td style="width:92px;">Zeiss</td>
			<td style="width:119px;"></td>
			<td>epifluorescence microscope</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:232px;">AxioCam MRm</td>
			<td style="width:92px;">Zeiss</td>
			<td style="width:119px;"></td>
			<td>CCD Camera</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:232px;">Tecnai TF20 Microscope</td>
			<td style="width:92px;">FEI</td>
			<td style="width:119px;"></td>
			<td>Transmisison Electron Miscroscope</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:232px;">TEM Eagle CCD</td>
			<td style="width:92px;">FEI</td>
			<td style="width:119px;"></td>
			<td>TEM CCD Camera</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:232px;">NanoBrook Omni DLS</td>
			<td style="width:92px;">Brookhaven</td>
			<td style="width:119px;"></td>
			<td style="width:283px;">Dynamic Light Scattering Instrument</td>
		</tr>
	</tbody>
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synthesis of cd-free inp/zns quantum dots suitable for biomedical applications

Fluorescent nanocrystals, specifically quantum dots, have been a useful tool for many biomedical applications. For successful use in biological systems, quantum dots should be highly fluorescent and small/monodisperse in size. While commonly used cadmium-based quantum dots possess these qualities, they are potentially toxic due to the possible release of Cd2+ ions through nanoparticle degradation. Indium-based quantum dots, specifically InP/ZnS, have recently been explored as a viable alternative to cadmium-based quantum dots due to their relatively similar fluorescence characteristics and size. The synthesis presented here uses standard hot-injection techniques for effective nanoparticle growth; however, nanoparticle properties such as size, emission wavelength, and emission intensity can drastically change due to small changes in the reaction conditions. Therefore, reaction conditions such temperature, reaction duration, and precursor concentration should be maintained precisely to yield reproducible products. Because quantum dots are not inherently soluble in aqueous solutions, they must also undergo surface modification to impart solubility in water. In this protocol, an amphiphilic polymer is used to interact with both hydrophobic ligands on the quantum dot surface and bulk solvent water molecules. Here, a detailed protocol is provided for the synthesis of highly fluorescent InP/ZnS quantum dots that are suitable for use in biomedical applications.

The uncoated InP cores do not demonstrate substantial visible fluorescence by eye. However, InP/ZnS (core/shell) quantum dots appear to fluoresce brightly by eye under UV irradiation. The fluorescence of InP/ZnS QDs was characterized using fluorescence spectroscopy. The fluorescence spectrum of QDs in hexanes (Figure 1) excited at 533 nm demonstrates one major peak centered at 600 nm with a full width at half maximum (FWHM) of 73 nm. While absorbance (0.2) offset in <stro...

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Synthesis of Cd-free InP/ZnS Quantum Dots Suitable for Biomedical Applications

In this protocol, the synthesis of Cd-free InP/ZnS quantum dots (QDs) is detailed. InP-based QDs are gaining popularity due to the toxicity of Cd2+ ions that may be released through nanoparticle degradation. After synthesis, QDs are solubilized in water using an amphiphilic polymer for use in biomedical applications.

Fluorescent nanocrystals, specifically quantum dots, have been a useful tool for many biomedical applications. For successful use in biological systems, ...

The overall goal of this procedure is to Synthesize Highly Fluorescent Indium Phosphide Zinc Sulfide Quantum Dots that are suitable for Biomedical Applications. This method can help clarify key questions about the Synthesis of Quantum Dot Nanoparticles by demonstrating specific steps that are crucial to achieve high quality Quantum Dots. The main advantage of this technique is the production of high quality, water soluble Quantum Dots.Viable for biological systems that can be synthesized in less than one day. To begin synthesis, fit a 100 milliliter round-bottom, three-neck flask with a 12-inch condenser. 0.398 grams of Indium Chloride, 30 milliliters Oleylamine, or OLA, and 0.245 grams of Zinc Chloride.Stir while evacuating at room temperature using a vacuum for one hour. The solution should appear colorless with a white precipitate. Use a heating mantle with a Thermocouple and PID Temperature Controller to increase the temperature of the solution to 120 degrees celsius.Use of a heating mantle and PID increases the uniformity and reproducibility of the reaction product. Evacuate the solution under vacuum for 20 minutes to remove low boiling point impurities that may affect core growth. Under inert gas, reflux the solution and increase the temperature to 223 degrees celsius for 15 minutes.The Indium Chloride and Zinc Chloride completely dissolve resulting in a pale yellow solution. Allow the temperature to stabilize for 10 minutes. Next, purge a disposable three milliliter plastic syringe and 4-inch 22 gauge needle with Nitrogen gas.The injection of TDMAP is a critical step in this procedure as the injection can greatly affect the dispersity of the resulting Indium Phosphide Quantum Dots. Using the syringe, quickly deliver 0.5 milliliters of TDMAP to the Indium Chloride solution. The solution temperature decreases slightly and returns to 220 degrees celsius while the solution changes from transparent pale yellow to opaque black.After nine and a half minutes, remove the reaction flask from the heating mantle until the temperature decreases below 200 degrees celsius. To protect the integrity of the Indium Phosphide cores proceed directly to the Zinc Sulfide coating. Place the reaction flask containing the Indium Phosphide Quantum Dots on a heating mantle and stabilize the temperature at 200 degrees celsius.Slowly add 3.58 grams of DDT to the solution over the course of 15 seconds. After allowing the solution to react for one hour, remove the reaction flask from the heating mantle and allow the solution to cool for approximately 60 degrees celsius. Once the Indium Phosphide Zinc Sulfide solution reaches approximately 60 degrees celsius, add 10 milliliters of Hexanes.Then, transfer the entire solution of roughly 45 milliliters to a 50 milliliter polypropylene centrifuge tube. Centrifuge the sample to remove unreacted solid precursors. Carefully remove the Supernatant and add 200 milliliters of Acetone.Equally divide the resulting solution into four 15 milliliters centrifuge tubes. Then, centrifuge the solution to precipitate the Indium Phosphide Zinc Sulfide Quantum Dots. Decant the supernatant and dry the Quantum Dot pellet thoroughly with Nitrogen gas to remove Acetone.Then, we suspend the Quantum Dots in 20 milliliters of OLA using Sonication. Then, transfer the resuspended Quantum Dots to a 50 milliliter round-bottom three-neck flask containing 0.474 grams of Zinc Stearate and stir. Evacuate the solution under vacuum for 20 minutes at room temperature.Under Nitrogen gas, increase the temperature to 180 degrees celsius and allow the reaction to proceed for three hours. Upon completion of the reaction, remove the flask from heating mantle and allow the solution to cool to approximately 60 degrees celsius. Once the Indium Phosphide Zinc Sulfide solution reaches approximately 60 degrees celsius, add 20 milliliters of Hexanes.Then, transfer the sample to a 50 milliliter polypropylene centrifuge tube before centrifuging to remove unreacted Zinc Stearate. Following centrifugation, add 200 milliliters of Acetone to the Supernatant and centrifuge using the same procedure as before. After thoroughly drying the pellet with Nitrogen gas, to remove the Acetone, dissolve the Indium Phosphide Zinc Sulfide Quantum Dot pellet in 30 milliliters of Hexanes.Vortex and sonicate the solution briefly to ensure complete dispersion. Removing the excess ligands is a critical step in this procedure. Failure to do so will negatively impact the efficiency of transferring the Quantum Dots into aqueous solutions using PMAL-d Repeat these purification steps two more times to ensure thorough removal of excess organic ligands.Interactions between the Amphiphilic Polymer and the Quantum Dot can be compromised in the presence of excess ligands. Following purification, determine the size and concentration of the synthesized Indium Phosphide Zinc Sulfide Quantum Dots using UV-Visible Spectroscopy. Dilute a portion of the synthesized Indium Phosphide Zinc Sulfide Quantum Dots with Hexanes to obtain one milliliter of one micromolar Quantum Dots.In a centrifuge tube, transfer 0.25 milliliters of Indium Phosphide Zinc Sulfide Quantum Dots into each tube. Add one milliliter of Acetone or menthol to the centrifuge tube, and centrifuge. Carefully remove the supernatant and dissolve each precipitate in one milliliter of Tetrahydrofuran or THF.Next, transfer the Indium Phosphide Zinc Sulfide Quantum Dots dissolved in THF into a 100 milliliter round-bottom flask and dilute with 16 milliliters of THF. To reduce the number of aggregates in solution, sonicate the Quantum Dots for five to ten minutes. Then, dissolve 30 milligrams of PMAL-d and 10 milliliters of molecular grade water.Water about sonication or gentle stirring until the solution is translucent is efficent to completely dissolve the polymer. The use of Vortex or a vigorous stirring can produce many bubbles which hinders interaction of the polymer with the Quantum Dot. Next, add the 10 milliliter polymer solution to the 100 milliliter round-bottom flask containing Indium Phosphide Zinc Sulfide Quantum Dots and THF.Evaporate the THF from the Quantum Dot polymer solution using a Rotary Evaporator with the flask in an ice bath to facilitate the interaction between the polymer and Quantum Dot. Once the solution is evaporated to 10 milliliters, remove the flask from the Rotary Evaporator and add 30 milliliters of molecular grade water. Return the flask to the Rotary Evaporator and continue to evaporate to two milliliters.This final evaporation step may take many hours. Ensure the ice bath is maintained. Remove the water soluble Indium Phosphide Zinc Sulfide Quantum Dots from the round-bottom flask with a pipette.Filter the Quantum Dot solution into a five milliliter centrifuge tube using a three milliliter plastic syringe attached to a 0.1 Micron Nylon Syringe Filter. Place the Quantum Dots into a 20 thousandth molecular weight cutoff membrane dialysis unit and dialyze against 05 Molar Borate Buffer, PH 8.5, to remove excess polymer. Using a vacuum concentrator, concentrate the Quantum Dots and Borate Buffer to one milliliter.For storage, perch the solution with Nitrogen gas before sealing with Parafilm. The water soluble Indium Phosphide Zinc Sulfide Quantum Dots are stable for at least four months at four degrees celsius in the dark. Shown here is the absorbance and corrected Florescence Admission Spectra of Indium Phosphide Zinc Sulfide Quantum Dots and Hexanes excited at 533 nanometers showing a maximum absorbance at 600 nanometers and a full width at half maximum of 73 nanometers.Here a TEM, or Transmission Electron Microscopy Image, of the Indium Phosphide Zinc Sulfide Quantum Dots dissolved in water is shown. A particle size distribution historgram of the TEM results in an average diameter of 2.74 plus or minus 0.72 nanometers. Single Florescent Puncta Analysis details the presence of distinct on and off states through a blinking profile of Indium Phosphide Zinc Sulfide Quantum Dots in water.This histogram exhibits the bimodal distribution of Pixel Intensity from one Quantum Dot blinking profile. Here a graph depicts the viability of N2a cells after incubation with Indium Phosphide Zinc Sulfide Quantum Dots for 24 or 48 hours. With one nanomolar to 25 nanomolar Quantum Dots.Negligible toxicity is observed below five nanomolar. Once mastered, this techniques can be completed in 12 hours if it is performed properly. After watching this video, you should have a good understanding of how to synthesize Highly Fluorescent Quantum Dots for use in Biomedical applications.Note that Allylamine is a hazardous, corrosive organic compound. Precautions such as wearing googles, gloves and a lab coat should always be taken when performing this procedure.

synthesis-cd-free-inpzns-quantum-dots-suitable-for-biomedical

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13.5K ビュー.  Missouri State University. The overall goal of this procedure is to Synthesize Highly Fluorescent Indium Phosphide Zinc Sulfide Quantum Dots that are suitable for Biomedical Applications. This method can help clarify key questions about the Synthesis of Quantum Dot Nanoparticles by demonstrating specific steps that are crucial to achieve high quality Quantum Dots. The main advantage of this technique is the production of high quality, water soluble Quantum Dots.Viable for biological systems that can be synthesiz...

ビデオ: Synthesis of Cd-free InP/ZnS Quantum Dots Suitable for Biomedical Applications

Preparation of Hydrophobic Metal-Organic Frameworks via Plasma Enhanced Chemical Vapor Deposition of Perfluoroalkanes for the Removal of Ammonia

Plasma enhanced chemical vapor deposition (PECVD) of perfluoroalkanes has long been studied for tuning the wetting properties of surfaces. For high surface area microporous materials, such as metal-organic frameworks (MOFs), unique challenges present themselves for PECVD treatments. Herein the protocol for development of a MOF that was previously unstable to humid conditions is presented. The protocol describes the synthesis of Cu-BTC (also known as HKUST-1), the treatment of Cu-BTC with PECVD of perfluoroalkanes, the aging of materials under humid conditions, and the subsequent ammonia microbreakthrough experiments on milligram quantities of microporous materials. Cu-BTC has an extremely high surface area (~1,800 m2/g) when compared to most materials or surfaces that have been previously treated by PECVD methods. Parameters such as chamber pressure and treatment time are extremely important to ensure the perfluoroalkane plasma penetrates to and reacts with the inner MOF surfaces. Furthermore, the protocol for ammonia microbreakthrough experiments set forth here can be utilized for a variety of test gases and microporous materials.

Herein the procedures for plasma enhanced chemical vapor deposition of perfluoroalkanes on microporous materials such as metal-organic frameworks to enhance their stability and hydrophobicity are described. Furthermore, breakthrough testing of milligram quantities of samples is described in detail.

Plasma enhanced chemical vapor deposition (PECVD) of perfluoroalkanes has long been studied for tuning the wetting properties of surfaces. For high ...

Synthesis of Immunotargeted Magneto-plasmonic Nanoclusters

Magnetic and plasmonic properties combined in a single nanoparticle provide a synergy that is advantageous in a number of biomedical applications including contrast enhancement in novel magnetomotive imaging modalities, simultaneous capture&#160;and detection of circulating tumor cells (CTCs),&#160;and multimodal molecular imaging combined with photothermal therapy of cancer cells. These applications have stimulated significant interest in development of protocols for synthesis of magneto-plasmonic nanoparticles with optical absorbance in the near-infrared (NIR) region and a strong magnetic moment. Here, we present a novel protocol for synthesis of such hybrid nanoparticles that is based on an oil-in-water microemulsion method. The unique feature of the protocol described herein is synthesis of magneto-plasmonic nanoparticles of various sizes from primary blocks which also have magneto-plasmonic characteristics. This approach yields nanoparticles with a high density of magnetic and plasmonic functionalities which are uniformly distributed throughout the nanoparticle volume. The hybrid nanoparticles can be easily functionalized by attaching antibodies through the Fc moiety leaving the Fab portion that is responsible for antigen binding available for targeting.

Here, we describe a protocol for synthesis of magneto-plasmonic nanoparticles with a strong magnetic moment and a strong near-infrared (NIR) absorbance. The protocol also includes antibody conjugation to the nanoparticles through the Fc moiety for various biomedical applications which require molecular specific targeting.

Magnetic and plasmonic properties combined in a single nanoparticle provide a synergy that is advantageous in a number of biomedical applications ...

Fabrication and Testing of Photonic Thermometers

In recent years, a push for developing novel silicon photonic devices for telecommunications has generated a vast knowledge base that is now being leveraged for developing sophisticated photonic sensors. Silicon photonic sensors seek to exploit the strong confinement of light in nano-waveguides to transduce changes in physical state to changes in resonance frequency. In the case of thermometry, the thermo-optic coefficient, i.e., changes in refractive index due to temperature, causes the resonant frequency of the photonic device such as a Bragg grating to drift with temperature. We are developing a suite of photonic devices that leverage recent advances in telecom compatible light sources to fabricate cost-effective photonic temperature sensors, which can be deployed in a wide variety of settings ranging from controlled laboratory conditions, to the noisy environment of a factory floor or a residence. In this manuscript, we detail our protocol for the fabrication and testing of photonic thermometers.

We describe the process of fabrication and testing of photonic thermometers.

In recent years, a push for developing novel silicon photonic devices for telecommunications has generated a vast knowledge base that is now being ...

Synthesis of Core-shell Lanthanide-doped Upconversion Nanocrystals for Cellular Applications

Lanthanide-doped upconversion nanocrystals (UCNs) have attracted much attention in recent years based on their promising and controllable optical properties, which allow for the absorption of near-infrared (NIR) light and can subsequently convert it into multiplexed emissions that span over a broad range of regions from the UV to the visible to the NIR. This article presents detailed experimental procedures for high-temperature co-precipitation synthesis of core-shell UCNs that incorporate different lanthanide ions into nanocrystals for efficiently converting deep-tissue penetrable NIR excitation (808 nm) into a strong blue emission at 480 nm. By controlling the surface modification with biocompatible polymer (polyacrylic acid, PAA), the as-prepared UCNs acquires great solubility in buffer solutions. The hydrophilic nanocrystals are further functionalized with specific ligands (dibenzyl cyclooctyne, DBCO) for localization on the cell membrane. Upon NIR light (808 nm) irradiation, the upconverted blue emission can effectively activate the light-gated channel protein on the cell membrane and specifically regulate the cation (e.g., Ca2+) influx in the cytoplasm. This protocol provides a feasible methodology for the synthesis of core-shell lanthanide-doped UCNs and subsequent biocompatible surface modification for further cellular applications.

A protocol is presented for the synthesis of core-shell lanthanide-doped upconversion nanocrystals (UCNs) and their cellular applications for channel protein regulation upon near-infrared (NIR) light illumination.

Lanthanide-doped upconversion nanocrystals (UCNs) have attracted much attention in recent years based on their promising and controllable optical ...

Probing the Structure and Dynamics of Interfacial Water with Scanning Tunneling Microscopy and Spectroscopy

Water/solid interfaces are ubiquitous and play a key role in many environmental, biophysical, and technological processes. Resolving the internal structure and probing the hydrogen-bond (H-bond) dynamics of the water molecules adsorbed on solid surfaces are fundamental issues of water science, which remains a great challenge owing to the light mass and small size of hydrogen. Scanning tunneling microscopy (STM) is a promising tool for attacking these problems, thanks to its capabilities of sub-&#197;ngstr&#246;m spatial resolution, single-bond vibrational sensitivity, and atomic/molecular manipulation. The designed experimental system consists of a Cl-terminated tip and a sample fabricated by dosing water molecules in situ onto the Au(111)-supported NaCl(001) surfaces. The insulating NaCl films electronically decouple the water from the metal substrates, so the intrinsic frontier orbitals of water molecules are preserved. The Cl-tip facilitates the manipulation of the single water molecules, as well as gating the orbitals of water to the proximity of Fermi level (EF) via tip-water coupling. This paper outlines the detailed methods of submolecular resolution imaging, molecular/atomic manipulation, and single-bond vibrational spectroscopy of interfacial water. These studies open up a new route for investigating the H-bonded systems at the atomic scale.

Here, we present a protocol to investigate the structure and dynamics of interfacial water at the atomic scale, in terms of submolecular resolution imaging, molecular manipulation, and single-bond vibrational spectroscopy.

Water/solid interfaces are ubiquitous and play a key role in many environmental, biophysical, and technological processes. Resolving the internal ...

Synthesis of Substrate-Bound Au Nanowires Via an Active Surface Growth Mechanism

Advancing synthetic capabilities is important for the development of nanoscience and nanotechnology. The synthesis of nanowires has always been a challenge, as it requires asymmetric growth of symmetric crystals. Here, we report a distinctive synthesis of substrate-bound Au nanowires. This template-free synthesis employs thiolated ligands and substrate adsorption to achieve the continuous asymmetric deposition of Au in solution at ambient conditions. The thiolated ligand prevented the Au deposition on the exposed surface of the seeds, so the Au deposition only occurs at the interface between the Au seeds and the substrate. The side of the newly deposited Au nanowires is immediately covered with the thiolated ligand, while the bottom facing the substrate remains ligand-free and active for the next round of Au deposition. We further demonstrate that this Au nanowire growth can be induced on various substrates, and different thiolated ligands can be used to regulate the surface chemistry of the nanowires. The diameter of the nanowires can also be controlled with mixed ligands, in which another &#34;bad&#34; ligand could turn on the lateral growth. With the understanding of the mechanism, Au nanowire-based nanostructures can be designed and synthesized.

We report a solution-based method to synthesize substrate-bound Au nanowires. By tuning the molecular ligands used during the synthesis, the Au nanowires can be grown from various substrates with different surface properties. Au nanowire-based nanostructures can also be synthesized by adjusting the reaction parameters.

Advancing synthetic capabilities is important for the development of nanoscience and nanotechnology. The synthesis of nanowires has always been a ...

Escherichia coli-Based Cell-Free Protein Synthesis: Protocols for a robust, flexible, and accessible platform technology

Over the last 50 years, Cell-Free Protein Synthesis (CFPS) has emerged as a powerful technology to harness the transcriptional and translational capacity of cells within a test tube. By obviating the need to maintain the viability of the cell, and by eliminating the cellular barrier, CFPS has been foundational to emerging applications in biomanufacturing of traditionally challenging proteins, as well as applications in rapid prototyping for metabolic engineering, and functional genomics. Our methods for implementing an E. coli-based CFPS platform allow new users to access many of these applications. Here, we describe methods to prepare extract through the use of enriched media, baffled flasks, and a reproducible method of tunable sonication-based cell lysis. This extract can then be used for protein expression capable of producing 900 &#181;g/mL or more of super folder green fluorescent protein (sfGFP) in just 5 h from experimental setup to data analysis, given that appropriate reagent stocks have been prepared beforehand. The estimated startup cost of obtaining reagents is $4,500 which will sustain thousands of reactions at an estimated cost of $0.021 per &#181;g of protein produced or $0.019 per &#181;L of reaction. Additionally, the protein expression methods mirror the ease of the reaction setup seen in commercially available systems due to optimization of reagent pre-mixes, at a fraction of the cost. In order to enable the user to leverage the flexible nature of the CFPS platform for broad applications, we have identified a variety of aspects of the platform that can be tuned and optimized depending on the resources available and the protein expression outcomes desired.

Escherichia coli-Based Cell-Free Protein Synthesis: Protocols for a robust, flexible, and accessible platform technology

This protocol details the steps, costs, and equipment necessary to generate E. coli-based cell extracts and implement in vitro protein synthesis reactions within 4 days or less. To leverage the flexible nature of this platform for broad applications, we discuss reaction conditions that can be adapted and optimized.

Over the last 50 years, Cell-Free Protein Synthesis (CFPS) has emerged as a powerful technology to harness the transcriptional and translational capacity ...

Synthesis of In37P20(O2CR)51 Clusters and Their Conversion to InP Quantum Dots

This text presents a method for the synthesis of In37P20(O2C14H27)51 clusters and their conversion to indium phosphide quantum dots. The In37P20(O2CR)51 clusters have been observed as intermediates in the synthesis of InP quantum dots from molecular precursors (In(O2CR)3, HO2CR, and P(SiMe3)3) and may be isolated as a pure reagent for subsequent study and use as a single-source precursor. These clusters readily convert to crystalline and relatively monodisperse samples of quasi-spherical InP quantum dots when subjected to thermolysis conditions in the absence of additional precursors above 200 &#176;C. The optical properties, morphology, and structure of both the clusters and quantum dots are confirmed using UV-Vis spectroscopy, photoluminescence spectroscopy, transmission electron microscopy, and powder X-ray diffraction. The molecular symmetry of the clusters is additionally confirmed by solution-phase 31P NMR spectroscopy. This protocol demonstrates the preparation and isolation of atomically-precise InP clusters, and their reliable and scalable conversion to InP QDs.

Synthesis of In37P20O2CR51 Clusters and Their Conversion to InP Quantum Dots

A protocol for the synthesis of In37P20(O2C14H27)51 clusters and their conversion to indium phosphide quantum dots is presented.

This text presents a method for the synthesis of In37P20(O2C14H27)51 clusters and their conversion to indium phosphide quantum dots. The In37P20(O2CR)51 ...

Comparison of Two Different Synthesis Methods of Single Crystals of Superconducting Uranium Ditelluride

Single crystal specimens of the actinide compound uranium ditelluride, UTe2, are of great importance to the study and characterization of its dramatic unconventional superconductivity, believed to entail spin-triplet electron pairing. A variety in the superconducting properties of UTe2 reported in the literature indicates that discrepancies between synthesis methods yield crystals with different superconducting properties, including the absence of superconductivity entirely. This protocol describes a process to synthesize crystals that exhibit superconductivity via chemical vapor transport, which has consistently exhibited a superconducting critical temperature of 1.6 K and a double transition indicative of a multi-component order parameter. This is compared to a second protocol that is used to synthesize crystals via the molten metal flux growth technique, which produces samples that are not bulk superconductors. Differences in the crystal properties are revealed through a comparison of structural, chemical, and electronic property measurements, showing that the most dramatic disparity occurs in the low-temperature electrical resistance of the samples.

Here, we present a protocol to synthesize two types of UTe2 crystals: those exhibiting robust superconductivity, via chemical vapor transport synthesis, and those lacking superconductivity, via molten metal flux synthesis.

Single crystal specimens of the actinide compound uranium ditelluride, UTe2, are of great importance to the study and characterization of its dramatic ...

Construction of Cyclic Cell-Penetrating Peptides for Enhanced Penetration of Biological Barriers

Cancer has been a grand challenge in global health. However, the complex tumor microenvironment generally limits the access of therapeutics to deeper tumor cells, leading to tumor recurrence. To conquer the limited penetration of biological barriers, cell-penetrating peptides (CPPs) have been discovered with excellent membrane translocation ability and have emerged as useful molecular transporters for delivering various cargoes into cells. However, conventional linear CPPs generally show compromised proteolytic stability, which limits their permeability across biological barriers. Thus, the development of novel molecular transporters that can penetrate biological barriers and exhibit enhanced proteolytic stability is highly desired to promote drug delivery efficiency in biomedical applications. We have previously synthesized a panel of short cyclic CPPs with aromatic crosslinks, which exhibited superior permeability in cancer cells and tissues compared to their linear counterparts. Here, a concise protocol is described for the synthesis of the fluorescently labeled cyclic polyarginine R8 peptide and its linear counterpart, as well as key steps for investigating their cell permeability.

This protocol describes the synthesis of cyclic cell-penetrating peptides with aromatic cross-links and the evaluation of their permeability across biological barriers.

Cancer has been a grand challenge in global health. However, the complex tumor microenvironment generally limits the access of therapeutics to deeper ...