Name: production of metal nanoparticles by pulsed laser-ablation in liquids: a tool for studying the antibacterial properties of nanoparticles
Uploaded: 2017-06-02T13:00:00
Description: Watch this Scientific Journal Video about Production of Metal Nanoparticles by Pulsed Laser-ablation in Liquids: A Tool for Studying the Antibacterial Properties of Nanoparticles at JoVE.com

Dette arbejde blev støttet af National Science Foundation (NSF tildeler CMMI-0922946 til DB, CMMI-1300920 til DB og S.O.M, og CMMI-1531789 til S.O&#39;M., DB og EAK) og en Busch Biomedicinsk Forskning Bevilgning til EAK og S.O&#39;M.

1. Fokusering af Nanosecond Laser og Measurement Fluence <ol><li> Saml ablationapparatet ved at placere en magnetisk omrøringsstang og et porøst ablationsstadium inde i et 50 ml glasbæger. BEMÆRK: Ablationsfasen består af en 3,81 cm diameter, 1,6 mm tykk rustfrit stålplatform med huller på 0,65 cm diameter og seks huller på 0,50 cm diameter boret i mønsteret illustreret i figur 1 . Formålet med disse huller er at tillade væsken at bevæge sig over målet...

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Reproducerbare antimikrobielle virkninger af NP&#39;er kræver konsekvent produktion af NP&#39;er med tilsvarende størrelser og koncentrationer. Derfor er det kritisk at standardisere laserparametre, herunder fluens, bølgelængde og pulsvarighed. Mens dynamisk lysspredning er en nem og hurtig metode til estimering af NP-størrelse, kræver præcis kvantificering af størrelsesfordelingen direkte måling af TEM. Da hver laserstråle har forskellige karakteristika med hensyn til tilstandsprofil og divergens, er det afg?...

Nanopartikler (NP&#39;er) defineres generelt som partikler, der har mindst en dimension, der er mindre end 100 nm i længden. Traditionelle kemiske NP syntese metoder kræver typisk farlige reduktionsmidler, såsom borhydrider og hydraziner. I modsætning hertil er laserablation af faste metalmål nedsænket i et flydende medium (pulserende laserablation i væsker - PLAL) en miljøvenlig rute for NP-syntese, som opfylder alle 12 af Green Chemistry 1 , 2 principperne. I PLAL bestråles et neddykket metalmål ved gentagne laserimpulser. Når laseren ablater målet, frigives en tæt plume af atomklynger og damp i det flydende medium, hvor NP&#39;er hurtigt koaleserer. NP&#39;er produceret af PLAL er fint dispergeret i et vandigt medium, og størrelsen, polydispersiteten og sammensætningen af ​​NP&#39;erne kan let styres ved at variere den vandige ablationsvæske såvel som laserparAmeter 1 , 2 , 3 , 4 , 5 , 6 . Nanopartikelegenskaber kan indstilles ved at justere et antal laserparametre, herunder: fluens, bølgelængde og pulsvarighed (gennemgået i reference 7 ). Laserfluens beregnes som pulsenergien divideret med området af laserplet på målfladen. De nøjagtige virkninger af fluens på størrelsen og polydispersiteten af ​​NP er noget kontroversielle. Generelt har det vist sig, at der for &#39;lange&#39; og &#39;ultrakort&#39; pulserede lasersystemer er lave og høje fluensregimer, der producerer negative og positive tendenser i størrelse henholdsvis 8 , 9 , 10 , 11 . NP størrelse distributionOns kan måles empirisk ved hjælp af teknikker som dynamisk lysspredning og transmissionselektronmikroskopi (TEM) som beskrevet nedenfor. Valget af laserbølgelængde kan påvirke de fysiske mekanismer, hvormed NP&#39;erne dannes. Ved kortere (ultraviolette) bølgelængder kan høj energi fotoner bryde interatomiske bindinger 12 . Denne mekanisme for fotoablation er et eksempel på en top-down NP-syntese, fordi den resulterer i frigivelsen af ​​ultra små fragmenter af materiale, der har tendens til at fremstille større mere polydisperse prøver ved quenching i nedsænkningsvæsken 12 , 13 , 14 . I modsætning hertil giver nær-infrarød ablation (A = 1,064 nm) en bottom-up-syntesemekanisme domineret af plasmaablation 12 . Laserabsorption ved målet frigør elektroner, der kolliderer med og efterfølgende fri, bundne elektroner. Som cOllisions stiger, materialet ioniseres og dermed antænder et plasma. Den omgivende væske begrænser plasmaet, forbedrer dets stabilitet og øger yderligere absorptionen 12 . Når det ekspanderende plasma blokeres af den begrænsende væske, kondenseres NP med forskellige geometrier 4 , 12 , 15 . Valget af laserpulsvarighed kan yderligere påvirke NP-dannelsesprocessen. Almindeligt anvendte langpulserede lasere med pulsvarigheder større end et par picosekunder omfatter alle milli-, mikro-, nano- og nogle picosecond-pulserende lasere. I dette pulsbredderegime er laserpulsvarigheden længere end elektronfonon-ækvilibreringstiden, som typisk er i rækkefølge af nogle få picosekunder 4 , 16 , 17 , 18 , 19. Dette resulterer i udslip af energi i det omgivende ablationsmedium og dannelsen af ​​NP&#39;er ved termiske mekanismer såsom termionisk emission, fordampning, kogning og smeltning 1 , 20 . Den antibakterielle aktivitet af NP&#39;er påvirkes stærkt af partikelstørrelse 21 , 22 , 23 , 24 . For at forbedre størrelsesreduktion og monodispersitet kan NP&#39;erne bestråles en anden gang ved anvendelse af en laser med en bølgelængde nær NP&#39;s overfladeplasmonresonans (SPR). Den indfaldende laserstråling absorberes af NP&#39;en gennem excitation af SPR. Fragmentering af NP kan forekomme ved enten termisk fordampning 25 , 26 eller Coulomb eksplosion 27 , 28 . Fotoexcitationen hæver tHan temperaturen af ​​NP over smeltepunktet, hvilket resulterer i afstivning af det ydre lag af partiklen. Det har vist sig, at tilsætningsmidler såsom polyvinylpyrrolidon (PVP) eller natriumdodecylsulfat (SDS) til opløsningen i høj grad kan forbedre efterbestrålingseffekter 5 . Virkningen af ​​tilsætningen af ​​forskellige opløste stoffer er blevet beskrevet i flere rapporter 1 , 4 , 6 . Den nemme manipulation af NP-egenskaber ved PLAL giver en ny metode til udvikling af nye NP-baserede antimikrobielle stoffer.

<table border="0" cellpadding="0" cellspacing="0" width="851">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:269px;">Nanosecond Nd:YAG laser</td>
			<td style="width:184px;">Ekspla</td>
			<td style="width:119px;">NL303</td>
			<td style="width:279px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:269px;">Motorized xy scanning stage</td>
			<td style="width:184px;">Standa</td>
			<td style="width:119px;">8MTF</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:269px;">UV-VIS spectrophotometer</td>
			<td style="width:184px;">Agilent</td>
			<td style="width:119px;">Cary 60</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:269px;">Dynamic light scattering unit</td>
			<td style="width:184px;">Malvern</td>
			<td style="width:119px;">Zetasizer ZS 90</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:269px;">Microbalance</td>
			<td style="width:184px;">Maktek</td>
			<td style="width:119px;">TM 400</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:269px;">Transmission electron microscope</td>
			<td style="width:184px;">Zeiss</td>
			<td style="width:119px;">EM 902</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:269px;">Silver foil target</td>
			<td style="width:184px;">Alfa Aesar</td>
			<td align="right" style="width:119px;">12127</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:269px;">250 mm focal length lens</td>
			<td style="width:184px;">Edmund Optics</td>
			<td style="width:119px;">69-624</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:269px;">Copper TEM grids</td>
			<td style="width:184px;">Pacific Grid-Tech</td>
			<td style="width:119px;">Cu-400LD</td>
			<td>Lacey/thin film coated grid</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:269px;">E. coli MG1655</td>
			<td style="width:184px;">ATCC</td>
			<td align="right" style="width:119px;">47076</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:269px;">Bacto-tryptone</td>
			<td style="width:184px;">BD Biosciences</td>
			<td align="right" style="width:119px;">211705</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:269px;">Yeast extract</td>
			<td style="width:184px;">BD Biosciences</td>
			<td align="right" style="width:119px;">212750</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:269px;">Sodium chloride</td>
			<td style="width:184px;">Fisher Scientific</td>
			<td style="width:119px;">BP3581</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:269px;">Bacto-agar</td>
			<td style="width:184px;">BD Biosciences</td>
			<td align="right" style="width:119px;">214010</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:269px;">Sodium dodecyl sulfate</td>
			<td style="width:184px;">Fisher Scientific</td>
			<td>BP166-100</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:269px;">Polyvinylpyrrolidone</td>
			<td style="width:184px;">Fisher Scientific</td>
			<td style="width:119px;">BP431-100</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:269px;">Stainless steel disc (for ablation stage)</td>
			<td style="width:184px;">Metal Remnants, Inc.</td>
			<td style="width:119px;">N/A</td>
			<td>1.5 inch diameter, 16 gauge</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:269px;">Beaker</td>
			<td style="width:184px;">Fisher Scientific</td>
			<td style="width:119px;">02-540G</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:269px;">Magnetic stir bar</td>
			<td style="width:184px;">Fisher Scientific</td>
			<td style="width:119px;">14-513-57</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:269px;">Magnetic stir plate</td>
			<td style="width:184px;">Fisher Scientific</td>
			<td style="width:119px;">11-100-49SH</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:269px;">Laser energy and power meter</td>
			<td style="width:184px;">Coherent</td>
			<td style="width:119px;">1098579</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:269px;">Carbon tape</td>
			<td style="width:184px;">Shinto Chemitron Co. Ltd.</td>
			<td style="width:119px;">STR Tape</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:269px;">Sonicating water bath</td>
			<td style="width:184px;">Branson</td>
			<td align="right" style="width:119px;">1510</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:269px;">Air compressor</td>
			<td style="width:184px;">GMC</td>
			<td style="width:119px;">Syclone 3010</td>
			<td>For drying ablation target</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:269px;">75 mm focal length lens</td>
			<td style="width:184px;">Edmund Optics</td>
			<td style="width:119px;">34-096</td>
			<td>Focusing lens for post-irradiation</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:269px;">Quartz cuvette</td>
			<td style="width:184px;">Precision Cells Inc</td>
			<td style="width:119px;">21UV40</td>
			<td>50 mm light path (for post-irradiation)</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:269px;">Kanamycin</td>
			<td style="width:184px;">Fisher Scientific</td>
			<td>BP906-5</td>
			<td></td>
		</tr>
		<tr height="105">
			<td height="105" style="height:105px;width:269px;">Light microscope</td>
			<td style="width:184px;">Nikon</td>
			<td style="width:119px;">50i</td>
			<td style="width:279px;">This microscope is used to focus the laser on the ablation stage. This particular model is no longer available, but any light microscope with a 4X objective will work.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:269px;">CCD camera</td>
			<td style="width:184px;">AmScope</td>
			<td style="width:119px;">MT5000-CCD</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:269px;">Micrometer slide</td>
			<td style="width:184px;">Ted Pella</td>
			<td style="width:119px;">2280-70</td>
			<td></td>
		</tr>
	</tbody>
</table>

production of metal nanoparticles by pulsed laser-ablation in liquids: a tool for studying the antibacterial properties of nanoparticles

Fremkomsten af ​​multidrugresistente bakterier er en global klinisk bekymring, der får nogle til at spekulere om vores tilbagevenden til en &quot;pre-antibiotics&quot; æra af medicin. Ud over bestræbelserne på at identificere nye antimikrobielle lægemidler med lille molekyl har der været stor interesse for brugen af ​​metal nanopartikler som belægninger til medicinsk udstyr, sårforbindelser og forbrugeremballage på grund af deres antimikrobielle egenskaber. Det store udvalg af metoder til nanopartikelsyntese resulterer i et bredt spektrum af kemiske og fysiske egenskaber, som kan påvirke antibakteriel virkning. Dette manuskript beskriver den pulserende laserablation i væsker (PLAL) metode til at skabe nanopartikler. Denne fremgangsmåde muliggør finjustering af nanopartikelstørrelse, sammensætning og stabilitet ved anvendelse af efterbestrålingsmetoder såvel som tilsætning af overfladeaktive stoffer eller volumenudslipningsmidler. Ved at kontrollere partikelstørrelse og sammensætning er der et stort antal fysiske og kemiske egenskaber ved metal nanopaRtikler kan udforskes, som kan bidrage til deres antimikrobielle effekt og derved åbne nye veje til antibakteriel udvikling.

Ved anvendelse af sølvmål fremstilles de ovenfor beskrevne laserparametre og 60 mM SDS i ablationsvæsken, sølv NP&#39;er med den karakteristiske UV-VIS-absorbans ved SPR ( figur 2A ). TEM og DLS målinger afslører en gennemsnitlig NP diameter på ca. 25 nm før efterbestråling ( Figur 2B ). Ablation af sølvmålet i 30 minutter giver typisk en NP-koncentration på 200 μg / ml. Ved vurdering af den antimikrobielle toksicit...

Watch this Scientific Journal Video about Production of Metal Nanoparticles by Pulsed Laser-ablation in Liquids: A Tool for Studying the Antibacterial Properties of Nanoparticles at JoVE.com

Production of Metal Nanoparticles by Pulsed Laser-ablation in Liquids: A Tool for Studying the Antibacterial Properties of Nanoparticles

De antimikrobielle egenskaber af metaller som kobber og sølv er blevet anerkendt i århundreder. Denne protokol beskriver pulserende laserablation i væsker, en metode til syntetisering af metal nanopartikler, der giver mulighed for at finjustere egenskaberne af disse nanopartikler for at optimere deres antimikrobielle virkninger.

Fremstilling af metal nanopartikler ved pulserende laserablation i væsker: Et værktøj til at studere de antibakterielle egenskaber ved nanopartikler

The emergence of multidrug-resistant bacteria is a global clinical concern leading some to speculate about our return to a "pre-antibiotics" era of ...

Research

JoVE Journal

Bioengineering

Fluorescent Nanoparticles for the Measurement of Ion Concentration in Biological Systems

Tightly regulated ion homeostasis throughout the body is necessary for the prevention of such debilitating states as dehydration.1 In contrast, rapid ion ...

Viral Nanoparticles for In vivo Tumor Imaging

Viral Nanoparticles for In vivo Tumor Imaging

Virale Nanopartikler for<em&gt; In vivo</em&gt; Tumor Imaging

The use of nanomaterials has the potential to revolutionize materials science and medicine. Currently, a number of different nanoparticles are being ...

Harmonic Nanoparticles for Regenerative Research

Harmonisk Nanopartikler for regenerativ Research

In this visualized experiment, protocol details are provided for in vitro labeling of human embryonic stem cells (hESC) with second harmonic generation ...

Biofunctionalized Prussian Blue Nanoparticles for Multimodal Molecular Imaging Applications

Biofunctionalized Prussian Blue Nanopartikler til multimodal Molekylære billedbehandling

Multimodal, molecular imaging allows the visualization of biological processes at cellular, subcellular, and molecular-level resolutions using multiple, ...

In Situ Mapping of the Mechanical Properties of Biofilms by Particle-tracking Microrheology

In Situ Mapping of the Mechanical Properties of Biofilms by Particle-tracking Microrheology

In Situ Kortlægning af de mekaniske egenskaber af biofilm fra Particle-tracking Microrheology

Bacterial cells are able to form surface-attached biofilm communities known as biofilms by encasing themselves in extracellular polymeric substances ...

Solid Lipid Nanoparticles (SLNs) for Intracellular Targeting Applications

Solid Lipid Nanoparticles SLNs for Intracellular Targeting Applications

Solid lipidnanopartikler (SLNs) til intracellulær målretning Applications

Nanoparticle-based delivery vehicles have shown great promise for intracellular targeting applications, providing a mechanism to specifically alter ...

Identification of Metal Oxide Nanoparticles in Histological Samples by Enhanced Darkfield Microscopy and Hyperspectral Mapping

Identifikation af Metal Oxide Nanopartikler i Histologiske prøver af Enhanced Darkfield mikroskopi og hyperspektral Mapping

Nanomaterials are increasingly prevalent throughout industry, manufacturing, and biomedical research. The need for tools and techniques that aid in the ...

Encapsulating Cytochrome c in Silica Aerogel Nanoarchitectures without Metal Nanoparticles while Retaining Gas-phase Bioactivity

Encapsulating Cytochrome c in Silica Aerogel Nanoarchitectures without Metal Nanoparticles while Retaining Gas-phase Bioactivity

indkapsling cytochrom<em&gt; c</em&gt; I Silica Aerogel Nanoarchitectures uden Metal Nanopartikler samtidig bevare Gas-fase Bioaktivitet

Applications such as sensors, batteries, and fuel cells have been improved through the use of highly porous aerogels when functional compounds are ...

Synthesis of Multi-walled Carbon Nanotubes Modified with Silver Nanoparticles and Evaluation of Their Antibacterial Activities and Cytotoxic Properties

Syntese af Multi-bjergomgivne kulstofnanorør modificeret med sølv nanopartikler og evaluering af deres antibakterielle aktiviteter og cytotoksiske egenskaber

In this study, multi-walled carbon nanotubes (MWCNTs) were treated with an aqueous sulfuric acid solution to form an oxygen-based functional group. Silver ...

Catalytic Scavenging of Plant Reactive Oxygen Species In Vivo by Anionic Cerium Oxide Nanoparticles

Catalytic Scavenging of Plant Reactive Oxygen Species In Vivo by Anionic Cerium Oxide Nanoparticles

Katalytisk Scavenging plante reaktive ilt arter In Vivo af anioniske Cerium oxid nanopartikler

Reactive oxygen species (ROS) accumulation is a hallmark of plant abiotic stress response. ROS play&#160;a dual role in plants by acting as signaling ...