Name: imaging of biological tissues by desorption electrospray ionization mass spectrometry
Uploaded: 2013-07-12T12:00:00
Description: Watch this Scientific Journal Video about Imaging of Biological Tissues by Desorption Electrospray Ionization Mass Spectrometry at JoVE.com

Dette arbeidet støttes av Arra NSF MR Instrument Development stipend # 0923179 til FMF. Vi takker Aqua Asberry, Lab koordinator for Parker H. Petit Institutt for bioteknologi og biovitenskap histologi Core for å få hjelp med vev snitting.

En. Tissue Seksjonering <ol><li> Store flash-frossen, hel vev i -80 ° C fryser til alt er klart for seksjonering. </li><li> Tillat den vevsprøve for å nå temperaturen i cryomicrotome før snitting (30 min). Sett kniven og prøven temperaturen til -30 ° C. </li><li> Når vevet har nådd den temperatur, håndtering av prøven med pinsett, skjære-eller baksiden av hjerne avhengig av hvilken del av hjernen som er av interesse for tilstrekkelig monteringsoverflaten (dvs. hvis fronten av hjernen er av primær...

<ol>
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G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Molecular+imaging+of+adrenal+gland+by+desorption+electrospray+ionization+mass+spectrometry.">Molecular imaging of adrenal gland by desorption electrospray ionization mass spectrometry.</a> Analyst. 135, 28-32 (2010).</li><li>Wiseman, J. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Desorption+electrospray+ionization+mass+spectrometry:+Imaging+drugs+and+metabolites+in+tissues.">Desorption electrospray ionization mass spectrometry: Imaging drugs and metabolites in tissues.</a> Proc. Natl. Acad. Sci. 105, 18120-18125 (2008).</li><li>Eberlin, L. S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Classifying+Human+Brain+Tumors+by+Lipid+Imaging+with+Mass+Spectrometry.">Classifying Human Brain Tumors by Lipid Imaging with Mass Spectrometry.</a> Cancer Res. 72, 645-654 (2012).</li><li>Wiseman, J. M., Ifa, D. R., Venter, A., Cooks, R. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ambient+molecular+imaging+by+desorption+electrospray+ionization+mass+spectrometry.">Ambient molecular imaging by desorption electrospray ionization mass spectrometry.</a> Nat. Protocols. 3, 517-524 (2008).</li><li>Van Berkel, G. J., Ford, M. J., Deibel, M. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Thin-Layer+Chromatography+and+Mass+Spectrometry+Coupled+Using+Desorption+Electrospray+Ionization.">Thin-Layer Chromatography and Mass Spectrometry Coupled Using Desorption Electrospray Ionization.</a> Anal. Chem. 77, 1207-1215 (2005).</li><li>Lane, A. L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Desorption+electrospray+ionization+mass+spectrometry+reveals+surface-mediated+antifungal+chemical+defense+of+a+tropical+seaweed.">Desorption electrospray ionization mass spectrometry reveals surface-mediated antifungal chemical defense of a tropical seaweed.</a> Proc. Natl. Acad. Sci. 106, 7314-7319 (2009).</li><li>Kertesz, V., Van Berkel, G. 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Optimalisering av DESI kilde geometri er avgjørende for vellykkede MSI eksperimenter. De mange variabler som bidrar til justering av systemet direkte påvirke følsomhet og oppløsning. Dersom det under optimalisering, har experimenter vanskeligheter med å få signal, anbefaler vi at du bruker rød tusj flekk trukket på lysbildet som en målestokk, fargestoff, rhodamine 6G, m / z 443, gir et sterkt signal i positiv ion modus, og kan brukes til initial optimalisering. I tillegg er løsemiddel seleksjon for Desi avgjø...

A correction was made to <a href="http://www.jove.com/video/50575" target="_parent">Imaging of Biological Tissues by Desorption Electrospray Ionization Mass Spectrometry</a>. There was an error with an author's name. The author's surname was appended with a missing character:
Rachel V. Bennett
instead of:
Rachel V. Bennet

Erratum: Imaging of Biological Tissues by Desorption Electrospray Ionization Mass Spectrometry

Vilkårlige bildebehandling ved massespektrometri forenkler oppkjøpet av kjemisk informasjon for oppdagelse og hypotese-genererende applikasjoner. Målsøkt billeddannende av en kjent kjemisk av interesse, på den annen side, kan lette økt følsomhet og selektivitet gjennom spesifikk metode utvikling. Massespektrometri imaging (MSI) er oftest utført på vev ved hjelp MALDI, en sekundær ion massespektrometri (SIMS), 2 og ambient ionisering teknikker, inkludert desorpsjon electrospray ionisering (DESI), 3 laser ablasjon-electrospray ionisering (LAESI), 4, 5 og væske mikro-junction-overflate prøvetakingssonde (LMJ-SSP). 6. I MALDI og SIMS, prøvene må fjernes fysisk fra prøven, og må være flat og tynn, slik de blir analysert ved høy-vakuum. MALDI krever belegg av prøven med en stråling-absorberende matrise, og legger til en ekstra og tungvint skritt til prøveopparbeidelse. SIMShar den høyeste lateral oppløsning, men bombardement med svært energiske partikler forårsaker omfattende molekylære fragmentering. Derfor MSI av omgivende metoder fylle en nisje hvor myk analyse med minimal prøveopparbeidelse er ønskelig. Men til dags dato, er alle metoder fremdeles begrenset av kravet om flate prøvenes overflater. Desi anvender en pneumatisk assistert ladet oppløsningsmiddel spray rettet mot sampeloverflaten å desorbere og ionisere analytter. Syv arbeidsmodell for desorpsjon og påfølgende ionisering av Desi er kjent som &quot;dråpe pick-up-modellen&quot;. 8-10 De ladede dråper primære produsert av desi sonde kolliderer med overflaten-, fukte-det og danner en tynn film i hvilken analytten er oppløst ved en faststoff-væske microextraction mekanisme 8. Etterfølgende dråpe kollisjoner resultat i impulsoverføringen og letting av sekundære dråper som inneholder det materialet ekstrahert fra overflaten . 9,10 syvende og gassfase ioner antas å bli produsert gjennom ESI-lignende prosesser etter ion fordampning, charge rester modeller eller andre modeller, 11 men den nøyaktige ion dannelsesprosessen i Desi ennå ikke er bevist eksperimentelt. 12. Desi følsomhet er sterkt avhengig av løseligheten av analytten i spray løsningsmiddel, som desorpsjon er avhengig av den lokaliserte microextraction. 13. Når den brukes med et programvarestyrt prøven stadium, prøven er skannet ensrettet med stepping kjørefelt under desi ionisering sonde, og gjennom den tid-domene, er m / z informasjon korreleres med det kjemiske stoff &#39;romlig fordeling (figur 1). Siden den første bevis på prinsippet DESI-MSI eksperiment rapportert av Van Berkel og Kertesz i 2006, har 14 teknikken modnet betraktelig, 15 med rapporterte søknader i analysen av lipider, 3,16 narkotika metabolitter, 17,18 diseaSE biomarkører, 19 hjernevev, 3,18,20 lungevev, 18 nyrevevet, 18 testikkel vev, 18 binyrene, 17 tynnskiktkromatografi plater, 21 og alger overflater. 22. Rutinen oppløsningen på bilder fremstilt ved DESI-MSI er 100-200 mikrometer, som er i siste instans bestemt av den effektive arealet trekkes ut av spray, men oppløsninger så lavt som 40 mikrometer er rapportert. 23-25 ​​Slike oppløsning og enkel analyse gjør DESI-MSI hensiktsmessig for rask og enkel analyse av biologiske vevsprøver med flater i 0,5-5 cm to utvalg, slik at anskaffelse av verdifull romlig informasjon for å forstå biologiske prosesser 26. Her, som et eksempel på en typisk DESI-MSI søknaden, vurderer vi de prosessuelle detaljene for å gjennomføre et vellykket eksperiment med avbildning av lipider i rotte hjernen vev. De to mest kritiske trinnene i protokollen, er detvevspreparat 27 og Desi ionekilde optimalisering, som beskrevet nedenfor.

<table border="1" fo:keep-together.within-page="always">
		<tbody>
		 <tr>
			<td>Reagents</td>
			<td></td>
			<td></td>
			<td></td>
		 </tr>
		 <tr>
			<td>Tissue-Tek O.C.T. Compound</td>
			<td>Sakura-Finetek</td>
			<td>4583</td>
			<td><a href="http://www.sakuraeu.com/products/showitem.asp?cat=11&amp;subcat=48" target="_blank">http://www.sakuraeu.com/products/showitem.asp?cat=11&amp;subcat=48</a></td>
		 </tr>
		 <tr>
			<td>Acetonitrile</td>
			<td>EMD</td>
			<td>AX0156-6</td>
			<td>OmniSolv, LC-MS Grade</td>
		 </tr>
		 <tr>
			<td>Acetic Acid</td>
			<td>Sigma Aldrich</td>
			<td>695092-500 ml</td>
			<td></td>
		 </tr>
		 <tr>
			<td>Equipment</td>
			<td></td>
			<td></td>
			<td></td>
		 </tr>
		 <tr>
			<td>Cryostat microtome</td>
			<td>Thermo Scientific</td>
			<td>CryoStar* NX70</td>
			<td>Any available microtome can be used for tissue sectioning <a href=" http://www.thermoscientific.com/ecomm/servlet/productsdetail?productId=13958375&amp;groupType=PRODUCT&amp;searchType=0&amp;storeId=11152&amp;from=search&amp;ca=cryostar" target="_blank"> http://www.thermoscientific.com/ecomm/servlet/productsdetail?productId=13958375&amp;groupType=PRODUCT&amp;searchType=0&amp;storeId=11152&amp;from=search&amp;ca=cryostar</a></td>
		 </tr>
		 <tr>
			<td>Omni Spray;DESI Spray Head</td>
			<td>Prosolia Inc.</td>
			<td></td>
			<td>Can also use the 2-D Omni Spray; Source kit instead of assembling components of imaging experiment <a href=" http://www.prosolia.com/sources.php" target="_blank"> http://www.prosolia.com/sources.php</a></td>
		 </tr>
		 <tr>
			<td>High Voltage Power Supply</td>
			<td>Stanford Research Systems, Inc.</td>
			<td>PS350/5000V-25W</td>
			<td><a href="http://www.thinksrs.com/products/PS300.htm" target="_blank">http://www.thinksrs.com/products/PS300.htm</a></td>
		 </tr>
		 <tr>
			<td>Rope heater, RTD, controller</td>
			<td>Omega</td>
			<td></td>
			<td><a href="http://www.omega.com/toc_asp/subsectionSC.asp?subsection=M02&amp;book=Heaters" target="_blank">http://www.omega.com/toc_asp/subsectionSC.asp?subsection=M02&amp;book=Heaters</a></td>
		 </tr>
		 <tr>
			<td>Labview</td>
			<td>National Instruments</td>
			<td>Version 7.1 </td>
			<td></td>
		 </tr>
		 <tr>
			<td>Translational stage</td>
			<td>Prior Scientific</td>
			<td>Optiscan II</td>
			<td><a href="http://www.prior.com/productinfo_auto_motorized_optiscan.html" target="_blank">http://www.prior.com/productinfo_auto_motorized_optiscan.html</a></td>
		 </tr>
		 <tr>
			<td>AccuTOF Mass Spectrometer</td>
			<td>JEOL</td>
			<td>JMS-T100LC</td>
			<td>Can use any mass spectrometer equipped with an extended capillary atmospheric pressure interface</td>
		 </tr>
		</tbody>
 </table>

imaging of biological tissues by desorption electrospray ionization mass spectrometry

Massespektrometri imaging (MSI) gir vilkårlige molekylær informasjon med høyest spesifisitet og romlig oppløsning for å undersøke biologiske vev på hundrevis til titalls mikrometer skala. Når arbeidet er utført under vanlige betingelser, blir prøven forbehandling unødvendig, og dermed forenkle den protokoll og samtidig opprettholde den høye kvaliteten på informasjon innhentet. Desorpsjon electrospray ionisering (DESI) er en spray-baserte ambient MSI teknikk som gir mulighet for direkte prøvetaking av overflater i friluft, selv in vivo. Når den brukes med et programvarestyrt prøven stadium, prøven rastered under desi ionisering sonde, og gjennom den tid-domene, er m / z informasjon korreleres med det kjemiske stoff &#39;romlig fordeling. Den riktighet av desi-MSI utgang er avhengig av kilde orientering og posisjonering i forhold til prøvens overflate og massespektrometer innløp. Heri, gjennomgår vi hvordan å forberede vevsdelene for DESI jegmaging og flere eksperimentelle forhold som direkte påvirker bildekvaliteten. Konkret beskriver vi protokollen for avbildning av rottehjernevev deler av DESI-MSI.

Figur 3 viser et representativt spektrum oppnådd fra en ubehandlet rotte hjerne seksjon. I den positive modus blir massespektrum dominert av fosfatidylcholiner grunn av sin høye effektivitet ionisering (tilskrives den positivt ladede kvaternær ammoniumgruppe). Den totale ion bildet av vevsdeler er også vist i figur 3, som viser rikelig signal på tvers av hele hjernen delen. Viktige lipider detekterte er identifisert i tabell 1 gjennom litteratur sammenligninger. </...

Watch this Scientific Journal Video about Imaging of Biological Tissues by Desorption Electrospray Ionization Mass Spectrometry at JoVE.com

Imaging of Biological Tissues by Desorption Electrospray Ionization Mass Spectrometry

Desorpsjon electrospray ionisering massespektrometri (DESI-MS) er en ambient metode der prøvene, inkludert biologisk vev, kan avbildes med minimal prøveopparbeidelse. Ved rastering prøven under ionisering sonde, gir denne spray-basert teknikk tilstrekkelig romlig oppløsning til å skjelne molekylære funksjoner av interesse innenfor vev seksjoner.

Avbildning av biologisk vev ved Desorpsjon Elektrospray Ionization massespektrometri

Mass spectrometry imaging (MSI) provides untargeted molecular information with the highest specificity and spatial resolution for investigating biological ...

Research

JoVE Journal

Bioengineering

Characterizing Bacterial Volatiles using Secondary Electrospray Ionization Mass Spectrometry (SESI-MS)

Characterizing Bacterial Volatiles using Secondary Electrospray Ionization Mass Spectrometry SESI-MS

Secondary electrospray ionization mass spectrometry (SESI-MS) is a method developed for the rapid detection of volatile compounds, without the need for ...

High Resolution 3D Imaging of Ex-Vivo Biological Samples by Micro CT

High Resolution 3D Imaging of Ex-Vivo Biological Samples by Micro CT

Non-destructive volume visualization can be achieved only by tomographic techniques, of which the most efficient is the x-ray micro computerized ...

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Cellular Lipid Extraction for Targeted Stable Isotope Dilution Liquid Chromatography-Mass Spectrometry Analysis

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The metabolism of fatty acids, such as arachidonic acid (AA) and linoleic acid (LA), results in the formation of oxidized bioactive lipids, including ...

Analyzing Cellular Internalization of Nanoparticles and Bacteria by Multi-spectral Imaging Flow Cytometry

Nanoparticulate  systems have emerged as valuable tools in vaccine delivery through their  ability to efficiently deliver cargo, including proteins, to ...

Sample Preparation Strategies for Mass Spectrometry Imaging of 3D Cell Culture Models

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Three dimensional cell cultures are attractive models for biological research. They combine the flexibility and cost-effectiveness of cell culture with ...

Qualitative Characterization of the Aqueous Fraction from Hydrothermal Liquefaction of Algae Using 2D Gas Chromatography with Time-of-flight Mass Spectrometry

Kvalitativ Karakterisering av den vandige fraksjon fra Hydrothermal kondensering av Alger Bruke 2D gasskromatografi med Time-of-flight massespektrometri

Two-dimensional gas chromatography coupled with time-of-flight mass spectrometry is a powerful tool for identifying and quantifying chemical components in ...

Whole-body Mass Spectrometry Imaging by Infrared Matrix-assisted Laser Desorption Electrospray Ionization (IR-MALDESI)

Whole-body Mass Spectrometry Imaging by Infrared Matrix-assisted Laser Desorption Electrospray Ionization IR-MALDESI

Hele kroppen massespektrometri Imaging av Infrared Matrix-assistert Laser desorpsjon electrospray ionisering (IR-MALDESI)

Ambient ionization sources for mass spectrometry (MS) have been the subject of much interest in the past decade. Matrix-assisted laser desorption ...

Engineering Three-dimensional Epithelial Tissues Embedded within Extracellular Matrix

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The architecture of branched organs such as the lungs, kidneys, and mammary glands arises through the developmental process of branching morphogenesis, ...

Engineering 'Golden' Fluorescence by Selective Pressure Incorporation of Non-canonical Amino Acids and Protein Analysis by Mass Spectrometry and Fluorescence