Name: methods to study lipid alterations in neutrophils and the subsequent formation of neutrophil extracellular traps
Uploaded: 2017-03-29T13:00:00
Description: Watch this Scientific Journal Video about Methods to Study Lipid Alterations in Neutrophils and the Subsequent Formation of Neutrophil Extracellular Traps at JoVE.com

Dette arbejde blev støttet af et fællesskab af Akademie für Tiergesundheit (Aid for Trade) og et stipendium fra ph.d.-studiet, &quot;Animal og zoonotiske infektioner,&quot; fra University of Veterinary Medicine, Hannover, Tyskland, forudsat at Ariane Neumann.

Indsamlingen af ​​det perifere blod i denne protokol blev godkendt af de lokale menneskelige forskningsetik provision. Alle mennesker, forudsat at deres skriftligt informeret samtykke. 1. Isolering af humant blod-afledte Neutrofiler ved densitetsgradientcentrifugering <ol><li> Isolering af humane blodafledte neutrofiler <ol><li> Lag ~ 20 ml blod på 20 ml af natrium diatrizoat / dextranopløsning nær en flamme og uden blanding. </li><li> Centrifugeres i 30 minutter ve...

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	<li>Riethmuller, J., Riehle, A., Grassme, H., Gulbins, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Membrane+rafts+in+host-pathogen+interactions.">Membrane rafts in host-pathogen interactions.</a> Biochim Biophys Acta. 1758 (12), 2139-2147 (2006).</li><li>Shahbazian, H., Atrian, A., Yazdanpanah, L., Lashkarara, G. R., Zafar Mohtashami, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Anti-inflammatory+effect+of+simvastatin+in+hemodialysis+patients.">Anti-inflammatory effect of simvastatin in hemodialysis patients.</a> Jundishapur J Nat Pharm Prod. 10, e17962 (2015).</li><li>Bavari, 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=Lipid+raft+microdomains:+A+gateway+for+compartmentalized+trafficking+of+Ebola+and+Marburg+viruses.">Lipid raft microdomains: A gateway for compartmentalized trafficking of Ebola and Marburg viruses.</a> J Exp Med. 195, 593-602 (2002).</li><li>Zaas, D. W., Duncan, M., Rae Wright, ., J, S. 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A., 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=Statins+Enhance+Formation+of+Phagocyte+Extracellular+Traps.">Statins Enhance Formation of Phagocyte Extracellular Traps.</a> Cell Host Microbe. 8 (5), 445-454 (2010).</li><li>Simons, K., Vaz, W. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Model+systems,+lipid+rafts,+and+cell+membranes.">Model systems, lipid rafts, and cell membranes.</a> Annu Rev Biophys Biomol Struct. 33, 269-295 (2004).</li><li>Bligh, E. G., Dyer, W. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+rapid+method+of+total+lipid+extraction+and+purification.">A rapid method of total lipid extraction and purification.</a> Can J Biochem Physiol. 37, 911-917 (1959).</li><li>Brogden, G., Propsting, M., Adamek, M., Naim, H. Y., Steinhagen, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Isolation+and+analysis+of+membrane+lipids+and+lipid+rafts+in+common+carp+(Cyprinus+carpio+L).">Isolation and analysis of membrane lipids and lipid rafts in common carp (Cyprinus carpio L).</a> Comp Biochem Physiol B Biochem Mol Biol. 169, 9-15 (2014).</li><li>Saldanha, T., Sawaya, A. C., Eberlin, M. 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V., 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=Lectin-induced+activation+of+plasma+membrane+NADPH+oxidase+in+cholesterol-depleted+human+neutrophils.">Lectin-induced activation of plasma membrane NADPH oxidase in cholesterol-depleted human neutrophils.</a> Arch Biochem Biophys. 516 (2), 173-181 (2011).</li><li>Masoud, R., Bizouarn, T., Hou&#233;e-Levin, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cholesterol:+A+modulator+of+the+phagocyte+NADPH+oxidase+activity+-+A+cell-free+study.">Cholesterol: A modulator of the phagocyte NADPH oxidase activity - A cell-free study.</a> Redox Biol. 3, 16-24 (2014).</li><li>Knight, J. 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=Peptidylarginine+deiminase+inhibition+is+immunomodulatory+and+vasculoprotective+in+murine+lupus.">Peptidylarginine deiminase inhibition is immunomodulatory and vasculoprotective in murine lupus.</a> J Clin Invest. 123 (7), 2981-2993 (2013).</li><li>Reichel, 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=Harbour+seal+(Phoca+vitulina)+PMN+and+monocytes+release+extracellular+traps+to+capture+the+apicomplexan+parasite+Toxoplasma+gondii.">Harbour seal (Phoca vitulina) PMN and monocytes release extracellular traps to capture the apicomplexan parasite Toxoplasma gondii.</a> Dev Comp Immunol. 50 (2), 106-115 (2015).</li><li>Chuammitri, P., 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=Chicken+heterophil+extracellular+traps+(HETs):+novel+defense+mechanism+of+chicken+heterophils.">Chicken heterophil extracellular traps (HETs): novel defense mechanism of chicken heterophils.</a> Vet Immunol Immunopathol. 129, 126-131 (2009).</li><li>Palic, D., Ostojic, J., Andreasen, C. 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De her beskrevne fremgangsmåder kan anvendes til at analysere specifikke lipider, såsom cholesterol, ved HPTLC eller HPLC og at undersøge virkningerne af farmakologiske Lipidforandringer på dannelsen af NET (se Neumann et al. 15). HPTLC er en relativt omkostningseffektiv og enkel metode til at analysere en lang række lipider i et stort antal prøver. Denne fremgangsmåde er blevet anvendt i mange forskningsområder, herunder antibiotisk kvantificering <s...

<html> Lipider er blevet vist at spille vigtige roller i cellehomøostase, celledød, værtspatogene interaktioner og cytokinfrigørelse 1. Over tid har interesse for og viden om virkningerne af lipider i vært-patogen interaktioner eller inflammation øges, og flere publikationer bekræfter den centrale rolle, som visse lipider, især steroid kolesterol, i cellulære responser. Farmakologisk behandling med statiner, som anvendes som inhibitorer af cholesterolbiosyntese ved at blokere 3-hydroxy-3-methylglutaryl-coenzym-A-reduktase (HMG-CoA-reduktase), kan fungere som anti-inflammatoriske midler ved at sænke serumniveauerne af interleukin 6 og C-reaktivt protein 2. Cholesterol og glycosphingolipid-berigede strukturer kan bruges af flere patogener, såsom bakterier og vira, som en gateway i værten 3, 4, 5,class = &quot;xref&quot;&gt; 6. Sphingolipider (fx sphingomyelin) er blevet vist at blive brugt af patogener til at fremme deres patogenicitet 7. I makrofager, mycobakterier brug cholesterolberiget domæner til indtastning celler; en udtynding af cholesterol hæmmer mycobakteriel optagelse 8. Endvidere infektion af makrofager med Francisella tularensis, en zoonotisk agens ansvarlig for tularemia (også kendt som kanin feber) 9, førte til en infektion, der blev afskaffet når cholesterol blev depleteret fra membranerne 10. Tilsvarende blev invasionen af værtsceller ved Escherichia coli via lipidrige strukturer vist sig at være kolesterol-afhængig 4. Desuden Salmonella typhimurium infektion eksperimenter af epitelceller viste, at cholesterol er afgørende for patogen indtrængen i cellerne 11. Cholesteroldepletering inhibited optagelsen af Salmonella 11. Endvidere en nylig undersøgelse fra gilk et al. viste, at kolesterol spiller en vigtig rolle i optagelsen af Coxiella burnetti 12. Derudover Tuong et al. konstateret, at 25-hydroxycholesterol spiller en afgørende rolle i fagocytose med lipopolysaccharid (LPS) -stimuleret makrofager 13. Fagocytose blev reduceret, når makrofager blev farmakologisk behandlet til nedbryder kolesterol 14. Således, cholesterol og andre lipider synes at spille en vigtig rolle i infektion og inflammation, idet deres udtømning kan reducere risikoen for invasion fra flere patogener 10, 11, 12. For nylig kunne vi vise, at lipid ombygninger, især udtømningen af ​​cholesterol fra cellen, inducere dannelsen af ​​neutrofil extracellular fælder (net) i humane blodafledte neutrofiler 15. Siden opdagelsen af NET i 2004, har de vist sig at spille kritiske roller i bakteriel indespærring, og dermed i at hindre spredning af smitte 16, 17. NET består af en DNA-rygrad forbundet med histoner, proteaser og antimikrobielle peptider 16. Frigivelse af NET ved neutrofiler kan induceres ved invaderende patogener 18, 19 og kemiske stoffer, såsom phorbol-myristat-acetat (PMA) eller statiner 16, 20. Imidlertid er de detaljerede cellulære mekanismer og især rollen af ​​lipider i denne proces, er stadig ikke helt klar. Analysen af ​​lipider kan føre til en bedre forståelse af de involverede i en lang række af cellulære processer og interaktioner, såsom frigivelse af NET mekanismer. Cholesterol og sphingomyelin er vitale bestanddele i cellemembranen og lipid mikrodomæner, hvor de tilføjer stabilitet og lette klyngedannelse af de involverede protein handel og signalhændelser 21-proteiner. For at undersøge den mekanistiske rolle, som visse lipider, amfifile farmakologiske midler, såsom det cykliske oligosaccharid methyl-β-cyclodextrin (MβCD), kan anvendes til at ændre lipidsammensætningen af en celle og til at reducere kolesterol in vitro 15. Her præsenteres en fremgangsmåde til at bruge HPTLC at analysere lipidsammensætningen af ​​neutrofiler i respons på MβCD. HPLC blev anvendt til at bekræfte niveauet af kolesterol i neutrofile befolkning. Desuden beskriver vi en metode til at visualisere dannelsen af ​​NET ved immunfluorescens mikroskopi i humane blodafledte neutrofiler i respons på MβCD.

<table>
	<tbody>
		<tr>
			<td>Neutrophil isolation, NET staining and quantification</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Alexa Flour 633 goat anti-rabbit IgG</td>
			<td>Invitrogen</td>
			<td>A-21070</td>
			<td></td>
		</tr>
		<tr>
			<td>Anti-MPO&#945; antibody</td>
			<td>Dako</td>
			<td>A0398</td>
			<td></td>
		</tr>
		<tr>
			<td>BSA</td>
			<td>Sigma-Aldrich</td>
			<td>3912-100G</td>
			<td></td>
		</tr>
		<tr>
			<td>Marienfeld-Neubauer improved counting chamber</td>
			<td>Celeromics</td>
			<td>MF-0640010</td>
			<td></td>
		</tr>
		<tr>
			<td>Confocal microscope TCS SP5 AOBS with tandem scanner</td>
			<td>Leica</td>
			<td>DMI6000CS</td>
			<td></td>
		</tr>
		<tr>
			<td>Dulbecco&#180;s PBS 10X</td>
			<td>Sigma-Aldrich</td>
			<td>P5493-1L</td>
			<td>Dilute 1:10 in water for 1X working solution</td>
		</tr>
		<tr>
			<td>Dy Light 488 conjugated highly cross-absorbed</td>
			<td>Thermo Fisher Scientific</td>
			<td>35503</td>
			<td></td>
		</tr>
		<tr>
			<td>Excel</td>
			<td>Microsoft</td>
			<td>2010</td>
			<td></td>
		</tr>
		<tr>
			<td>DNA/Histone 1 antibody</td>
			<td>Millipore</td>
			<td>MAB3864</td>
			<td></td>
		</tr>
		<tr>
			<td>Image J</td>
			<td>NIH</td>
			<td>1.8</td>
			<td>http://imagej.nih.gov/ij/</td>
		</tr>
		<tr>
			<td>Light microscope</td>
			<td>VWR</td>
			<td>630-1554</td>
			<td></td>
		</tr>
		<tr>
			<td>Methyl-&#946;-cyclodextrin</td>
			<td>Sigma-Aldrich</td>
			<td>C4555-1G</td>
			<td></td>
		</tr>
		<tr>
			<td>PFA</td>
			<td>Carl Roth</td>
			<td>0335.3</td>
			<td>dissolve in water, heat up to 65 &#176;C and add 1N NaOH to clear solution</td>
		</tr>
		<tr>
			<td>PMA</td>
			<td>Sigma-Aldrich</td>
			<td>P8139-1MG</td>
			<td>Stock 16 &#181;M, dissolved in 1X PBS</td>
		</tr>
		<tr>
			<td>Poly-L-lysine</td>
			<td>Sigma-Aldrich</td>
			<td>P4707</td>
			<td></td>
		</tr>
		<tr>
			<td>Polymorphprep</td>
			<td>AXIS-SHIELD</td>
			<td>AN1114683</td>
			<td></td>
		</tr>
		<tr>
			<td>ProLong Gold antifade reagent with DAPI</td>
			<td>Invitrogen</td>
			<td>P7481</td>
			<td></td>
		</tr>
		<tr>
			<td>Quant-iT PicoGreen dsDNA Reagent</td>
			<td>Invitrogen</td>
			<td>P7581</td>
			<td></td>
		</tr>
		<tr>
			<td>RPMI1640</td>
			<td>PAA</td>
			<td>E 15-848</td>
			<td></td>
		</tr>
		<tr>
			<td>HBSS with CaCl and Mg</td>
			<td>Sigma</td>
			<td>H6648</td>
			<td></td>
		</tr>
		<tr>
			<td>Triton X-100</td>
			<td>Sigma-Aldrich</td>
			<td>T8787-50ml</td>
			<td></td>
		</tr>
		<tr>
			<td>Trypanblue</td>
			<td>Invitrogen</td>
			<td>15250-061</td>
			<td>0.4% solution</td>
		</tr>
		<tr>
			<td>Water</td>
			<td>Carl Roth</td>
			<td>3255.1</td>
			<td>endotoxin-free</td>
		</tr>
		<tr>
			<td>Name</td>
			<td>Company</td>
			<td>Catalog Number&#160;</td>
			<td>Comments</td>
		</tr>
		<tr>
			<td>Lipid isolation and analysis</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>1-propanol</td>
			<td>Sigma-Aldrich</td>
			<td>33538</td>
			<td></td>
		</tr>
		<tr>
			<td>10 &#181;l syringe</td>
			<td>Hamilton</td>
			<td>701 NR 10 &#181;l</td>
			<td></td>
		</tr>
		<tr>
			<td>Diethyl ether</td>
			<td>Sigma-Aldrich</td>
			<td>346136</td>
			<td></td>
		</tr>
		<tr>
			<td>Ethyl acetate</td>
			<td>Carl Roth</td>
			<td>7336.2</td>
			<td></td>
		</tr>
		<tr>
			<td>Canullla 26G</td>
			<td>Braun</td>
			<td>4657683</td>
			<td></td>
		</tr>
		<tr>
			<td>Copper(II)sulphatepentahydrate</td>
			<td>Merck</td>
			<td>1027805000</td>
			<td></td>
		</tr>
		<tr>
			<td>Chloroform</td>
			<td>Carl Roth</td>
			<td>7331.1</td>
			<td></td>
		</tr>
		<tr>
			<td>CP ATLAS software</td>
			<td>Lazarsoftware</td>
			<td>2.0</td>
			<td></td>
		</tr>
		<tr>
			<td>Chromolith HighResolution RP-18 endcapped 100-4.6 mm column</td>
			<td>Merck</td>
			<td>152022</td>
			<td></td>
		</tr>
		<tr>
			<td>High Performance Liquid Chromatograph Chromaster</td>
			<td>Hitachi</td>
			<td>HITA 892-0080-30Y</td>
			<td>Paramaters are dependent on individual HPLC machine</td>
		</tr>
		<tr>
			<td>HPLC UV Detector</td>
			<td>Hitachi</td>
			<td>5410</td>
			<td></td>
		</tr>
		<tr>
			<td>HPLC Column Oven</td>
			<td>Hitachi</td>
			<td>5310</td>
			<td></td>
		</tr>
		<tr>
			<td>HPLC Auto Sampler</td>
			<td>Hitachi</td>
			<td>5260</td>
			<td></td>
		</tr>
		<tr>
			<td>HPLC Pump</td>
			<td>Hitachi</td>
			<td>5160</td>
			<td></td>
		</tr>
		<tr>
			<td>Methanol</td>
			<td>Carl Roth</td>
			<td>7342.1</td>
			<td></td>
		</tr>
		<tr>
			<td>n-Hexane</td>
			<td>Carl Roth</td>
			<td>7339.1</td>
			<td></td>
		</tr>
		<tr>
			<td>Phosphoric acid</td>
			<td>Sigma-Aldrich</td>
			<td>30417</td>
			<td></td>
		</tr>
		<tr>
			<td>Potassium chloride</td>
			<td>Merck</td>
			<td>49,361,000</td>
			<td></td>
		</tr>
		<tr>
			<td>Potters</td>
			<td>LAT Garbsen</td>
			<td>5 ml</td>
			<td></td>
		</tr>
		<tr>
			<td>SDS</td>
			<td>Carl Roth</td>
			<td>CN30.3</td>
			<td></td>
		</tr>
		<tr>
			<td>HPTLC silica gel 60</td>
			<td>Merck</td>
			<td>105553</td>
			<td></td>
		</tr>
		<tr>
			<td>Vacufuge plus basic device</td>
			<td>Eppendorf</td>
			<td>22820001</td>
			<td></td>
		</tr>
		<tr>
			<td>Corning Costar cell culture 48-well plate, flat bottom</td>
			<td>Sigma</td>
			<td>CLS3548</td>
			<td></td>
		</tr>
		<tr>
			<td>Coverslip</td>
			<td>Thermo Fisher Scientific</td>
			<td>1198882</td>
			<td></td>
		</tr>
		<tr>
			<td>Glass slide</td>
			<td>Carl Roth</td>
			<td>1879</td>
			<td></td>
		</tr>
		<tr>
			<td>BD Tuberculin Syringe Only 1 ml</td>
			<td>BD Bioscience</td>
			<td>309659</td>
			<td></td>
		</tr>
	</tbody>
</table>

methods to study lipid alterations in neutrophils and the subsequent formation of neutrophil extracellular traps

Lipid analyse udført ved højtydende tyndtlagskromatografi (HPTLC) er en relativt enkel, omkostningseffektiv fremgangsmåde til analyse af et bredt udvalg af lipider. Funktionen af lipider (fx i værtspatogene interaktioner eller vært post) er blevet rapporteret at spille en afgørende rolle i cellulære processer. Her viser vi en metode til at bestemme lipidsammensætning, med fokus på cholesterolniveauet af primære blodafledte neutrofiler ved HPTLC i sammenligning med højtryksvæskechromatografi (HPLC). Formålet var at undersøge rollen af ​​lipid / cholesterol ændringer i dannelsen af ​​neutrofile ekstracellulære fælder (NET). NET frigivelse er kendt som en værtsforsvarsmekanisme at forhindre patogener i at sprede i værten. Derfor blev afledt af blod humane neutrofiler behandlet med methyl-β-cyclodextrin (MβCD) for at inducere lipid forandringer i cellerne. Anvendelse HPTLC og HPLC, har vi vist, at MβCD behandling af cellerne fører til lipidændringer associeret med en signifikant reduktion i indholdet af cellen kolesterol. Samtidig, MβCD behandling af neutrofilerne førte til dannelsen af ​​NET, som vist ved immunfluorescens mikroskopi. Sammenfattende her præsenterer vi en detaljeret metode til at studere lipid ændringer i neutrofiler og dannelsen af ​​NET.

Humane blodafledte neutrofiler blev isoleret ved densitetsgradientcentrifugering (figur 2). For at undersøge virkningen af ​​lipid ændringer på neutrofiler, blev cellerne behandlet med 10 mM MβCD, som udtømmer cholesterol fra cellen. Efterfølgende blev lipiderne isoleres fra prøver ved Bligh og Dyer (figur 1, venstre panel), som beskrevet af Brogden et al. 23. De fremstillede lipid prøver blev fyldt på silica...

Watch this Scientific Journal Video about Methods to Study Lipid Alterations in Neutrophils and the Subsequent Formation of Neutrophil Extracellular Traps at JoVE.com

Methods to Study Lipid Alterations in Neutrophils and the Subsequent Formation of Neutrophil Extracellular Traps

Lipider er kendt for at spille en vigtig rolle i cellulære funktioner. Her beskriver vi en fremgangsmåde til bestemmelse lipidsammensætningen af ​​neutrofiler, med vægt på kolesterolniveauet, ved hjælp af både HPTLC og HPLC at få en bedre forståelse af de underliggende mekanismer for neutrofil ekstracellulær fælde dannelse.

Metoder til at studere Lipidforandringer i neutrofiler og den efterfølgende dannelse af neutrofile ekstracellulær fælder

Lipid analysis performed by high performance thin layer chromatography (HPTLC) is a relatively simple, cost-effective method of analyzing a broad range of ...

Research

JoVE Journal

Immunology and Infection

Neutrophil Extracellular Traps: How to Generate and Visualize Them

Neutrophil granulocytes are the most abundant group of leukocytes in the peripheral blood. As professional phagocytes, they engulf bacteria and kill them ...

Simplified Human Neutrophil Extracellular Traps (NETs) Isolation and Handling

Simplified Human Neutrophil Extracellular Traps NETs Isolation and Handling

Forenklet human neutrofil Ekstracellulære Fælder (NET) Isolering og Håndtering

Neutrophil Extracellular Traps (NETs) have been recently identified as part of the neutrophil&#8217;s antimicrobial armamentarium. Apart from their role ...

High Throughput Measurement of Extracellular DNA Release and Quantitative NET Formation in Human Neutrophils In Vitro

High Throughput Measurement of Extracellular DNA Release and Quantitative NET Formation in Human Neutrophils In Vitro

High Throughput Måling af ekstracellulær DNA udledning og kvantitativ NET Formation i humane neutrofiler<em&gt; In vitro</em

Neutrophil granulocytes are the most abundant leukocytes in the human blood. Neutrophils are the first to arrive at the site of infection. Neutrophils ...

In Vitro Canine Neutrophil Extracellular Trap Formation: Dynamic and Quantitative Analysis by Fluorescence Microscopy

In Vitro Canine Neutrophil Extracellular Trap Formation: Dynamic and Quantitative Analysis by Fluorescence Microscopy

In Vitro Canine neutrofile ekstracellulære fælde dannelse: Dynamisk og kvantitativ analyse af Fluorescens mikroskopi

In response to invading pathogens, neutrophils release neutrophil extracellular traps (NETs), which are extracellular networks of DNA decorated with ...

Automated Image-Based Quantification of Neutrophil Extracellular Traps Using NETQUANT

Automatiseret billedbaseret kvantificering af neutrofile ekstracellulære fælder med NETQUANT

Neutrophil extracellular traps (NETs) are web-like antimicrobial structures consisting of DNA and granule derived antimicrobial proteins. ...

A High-throughput Assay to Assess and Quantify Neutrophil Extracellular Trap Formation

En høj overførselshastighed analyse til at vurdere og kvantificere neutrofile ekstracellulære fælde dannelse

Neutrophil extracellular traps (NETs) are immunogenic extracellular DNA structures that can be released by neutrophils upon a wide variety of triggers. ...

Identification of Neutrophil Extracellular Traps in Paraffin-Embedded Feline Arterial Thrombi using Immunofluorescence Microscopy

Identifikation af neutrofile ekstracellulære fælder i paraffin-embedded feline arteriel trombose ved hjælp af Immunofluorescence Mikroskopi

Neutrophil extracellular traps (NETs), composed of cell-free DNA (cfDNA) and proteins like histones and neutrophil elastase (NE), are released by ...

Morphological and Compositional Analysis of Neutrophil Extracellular Traps Induced by Microbial and Chemical Stimuli

Morfologisk og sammensætningsanalyse af neutrofile ekstracellulære fælder induceret af mikrobielle og kemiske stimuli

Neutrophils function as the first line of cellular defense in an innate immune response by employing diverse mechanisms, such as the formation of ...

Quantifying Myeloperoxidase-DNA and Neutrophil Elastase-DNA Complexes from Neutrophil Extracellular Traps by Using a Modified Sandwich ELISA

Kvantificering af myeloperoxidase-DNA og neutrofile elastase-DNA-komplekser fra neutrofile ekstracellulære fælder ved hjælp af en modificeret sandwich-ELISA

Certain stimuli, such as microorganisms, cause neutrophils to release neutrophil extracellular traps (NETs), which are basically web-like structures ...

Staining and Imaging to Visualize Neutrophil Extracellular Traps

Immunofluorescence Imaging of Neutrophil Extracellular Traps in Human and Mouse Tissues

Neutrophil Extracellular Traps Imaging in Human and Mouse Tissues (Video) | JoVE

Immunofluorescensbilleddannelse af neutrofile ekstracellulære fælder i væv fra mennesker og mus

Neutrophil extracellular traps (NETs) are released by neutrophils as a response to bacterial infection or traumatic tissue damage but also play a role in ...