Name: synthesis of cd-free inp/zns quantum dots suitable for biomedical applications
Uploaded: 2016-02-06T14:00:00
Description: Watch this Scientific Journal Video about Synthesis of Cd-free InP/ZnS Quantum Dots Suitable for Biomedical Applications at JoVE.com

Os autores agradecem o Departamento de Química e da Faculdade de Pós-Graduação da Universidade Estadual do Missouri para o seu apoio a este projeto. Reconhecemos também o Electron Microscopy Laboratório do Laboratório Nacional Frederick para Pesquisa do Câncer para o uso de seu microscópio eletrônico de transmissão e redes revestidas de carbono.

1. Síntese de fosfeto de índio / Zinc Sulfide (INP / ZnS) Quantum Dots <ol><li> Síntese de fosfeto de índio (InP) Quantum Dot Cores <ol><li> Caber um fundo redondo de 100 ml, 3-neck, garrafa com um condensador de 12 polegadas. Adicionar 30 ml de oleilamina (OLA), cloreto de 0,398 g de índio (III) (incl 3), (II), cloreto de 0,245 g de zinco (ZnCl2) e agita-se durante a evacuação, à TA utilizando um vácuo durante 1 hora. A solução deve aparecer incolor com um precipitado...

<ol>
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Este protocolo detalha a síntese das altamente fluorescentes QDs InP / ZnS que podem ser utilizados em muitos sistemas biológicos. Os produtos sintetizados QD aqui exibiu um único pico de emissão de fluorescência centrada a 600 nm com um FWHM de 73 nm (Figura 1), que é comparável a outras sínteses descritas anteriormente 12. O tempo de reacção e temperatura da reacção são extremamente passos cruciais, devido ao seu profundo efeito sobre a qualidade de síntese QD e repetibilidade...

Os pontos quânticos (QDs) são semicondutor nanocristais que exibem propriedades fluorescentes quando irradiado com luz 1. Devido ao seu pequeno tamanho (2-5 nm), que é semelhante a muitas biomoléculas grandes, e facilidade de biofunctionalization, QDs são uma ferramenta extremamente atraente para aplicações biomédicas. Eles têm encontrado utilização na rotulagem biológica, imagem única molécula-célula viva, a entrega de drogas, imagem in vivo, detecção de patógenos e rastreamento de celular, entre muitos outros usos 2-8. QDs baseada em CD foram mais comumente usados ​​em aplicações biomédicas devido à sua intensa fluorescência e larguras de pico de emissão estreitas 9. No entanto, as preocupações foram levantadas devido ao potencial de toxicidade do Cd 2+ 10 que pode ser introduzida através da degradação das nanopartículas. Recentemente, QDs baseados em InP têm sido explorados como uma alternativa para QDs baseadas em CD porque eles mantêm muitas características de fluorescênciade QDs Cd-baseado e pode ser mais biocompatível 11. QDs baseadas em CD foram encontrados para ser significativamente mais tóxica do que QDs baseados em InP em ensaios in vitro para concentrações tão baixas quanto 10 pM, após apenas 48 h 11. A cor de emissão de fluorescência de QDs é tamanho-ajustável 1. Isto é, como o tamanho das QD aumenta, a emissão de fluorescência é vermelho-deslocado. O tamanho e dimensão dispersibilidade dos produtos QD pode ser modificada alterando a temperatura, duração da reacção, ou em condições de concentração de precursor durante a reacção 12. Embora o pico de emissão de InP QDs é tipicamente mais ampla e menos intensa do que QDs baseadas em CD, InP QDs pode ser feita numa grande variedade de cores, destinadas a evitar sobreposição espectral, e são suficientemente intensa para a maioria das aplicações biomédicas 12. A síntese detalhada neste protocolo produz QDs com um pico de emissão vermelha centrado em 600 nm. Várias medidas são tomadas afsíntese ter dos núcleos QD para manter a integridade óptica dos QDs e torná-los compatíveis para aplicações biológicas. A superfície do núcleo QD deve ser protegida contra a oxidação ou defeitos superficiais, que podem causar têmpera; portanto, uma concha ZnS é revestido sobre o núcleo para produzir InP / ZnS (core / shell) qds 13. Este revestimento tem sido demonstrado para proteger o fotoluminescência do produto QD. A presença de iões de zinco, durante a síntese InP QD foi mostrado para limitar os defeitos de superfície, bem como a distribuição do tamanho de redução 12. Mesmo com a presença de Zn2 + no meio de reacção, a síntese de InZnP são altamente improvável 12. Depois do revestimento, resultantes QDs InP / ZnS são revestidos em ligandos hidrofóbicos, tais como o óxido de trioctilfosfina (TOPO) ou oleilamina 12,14. Um polímero anfifílico pode interagir com ligandos hidrófobos na superfície do QD, bem como moléculas de água em massa para conferir solubilidade em água 15. polímeros anfifílicos com carbogrupos químicos xylate pode ser utilizado como &quot;alças&quot; químicas para funcionalizar ainda mais os QDs. Este protocolo detalha a síntese e funcionalização de solúveis em água InP / ZnS QDs com emissão de fluorescência muito intenso e relativamente pequeno em tamanho dispersity. Estes QDs são potencialmente menos tóxico do que QDs CdSe / ZnS vulgarmente utilizados. Nisto, a síntese de InP / ZnS QDs fornece uma alternativa prática para QDs baseadas em CD para aplicações biomédicas.

<table border="0" cellpadding="0" cellspacing="0" width="725">
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		<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>
</table>

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.

Os núcleos de InP não revestidos não demonstram fluorescência visível substancial a olho nu. No entanto, InP / ZnS (core / shell) pontos quânticos parecem apresentam forte fluorescência por olho sob irradiação UV. A fluorescência de InP / ZnS QDs foi caracterizado usando espectroscopia de fluorescência. O espectro de fluorescência de QDs em hexanos (Figura 1) excitado a 533 nm demonstra um pico principal centrado a 600 nm com uma largura total a meia altura (...

Watch this Scientific Journal Video about Synthesis of Cd-free InP/ZnS Quantum Dots Suitable for Biomedical Applications at JoVE.com

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.

Síntese de Cd-livre InP / ZnS Quantum Dots Indicado para aplicações biomédicas

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

Research

JoVE Journal

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