Name: assessing anti-fungal activity of isolated alveolar macrophages by confocal microscopy
Uploaded: 2014-07-09T21:30:00
Description: Watch this Scientific Journal Video about Assessing Anti-fungal Activity of Isolated Alveolar Macrophages by Confocal Microscopy at JoVE.com

Questo lavoro è stato sostenuto dal National Institute of Allergy and Infectious Disease di Grant R01AI079253 (a BHS), e dal National Cancer Institute Cancer Support Center Concessione CA016056 al Roswell Park Cancer Institute.

Tutte le procedure eseguite su animali in questo studio sono stati approvati dal Comitato cura degli animali ed uso al Roswell Park Cancer Institute e rispettate tutte le statali, federali e istituti nazionali di regolamentazione di salute. 1. In vivo macrofagi Labeling Nota: PHK26 è un colorante lipofilo che sarà ripreso da fagociti sia in circolazione e nei tessuti quando somministrato per via endovenosa (iv). PKH26 è stato utilizzato per l&#39;etiche...

<ol>
	<li>Reeves, E. 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=Killing+activity+of+neutrophils+is+mediated+through+activation+of+proteases+by+K++flux.">Killing activity of neutrophils is mediated through activation of proteases by K+ flux.</a> Nature. 416, 291-297 (2002).</li><li>Bianchi, 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=Restoration+of+NET+formation+by+gene+therapy+in+CGD+controls+aspergillosis.">Restoration of NET formation by gene therapy in CGD controls aspergillosis.</a> Blood. 114, 2619-2622 (2009).</li><li>Vethanayagam, R. R., 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=Role+of+NADPH+oxidase+versus+neutrophil+proteases+in+antimicrobial+host+defense.">Role of NADPH oxidase versus neutrophil proteases in antimicrobial host defense.</a> PLoS One. 6, (2011).</li><li>Grimm, M. J., 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=Monocyte-+and+Macrophage-Targeted+NADPH+Oxidase+Mediates+Antifungal+Host+Defense+and+Regulation+of+Acute+Inflammation+in+Mice.">Monocyte- and Macrophage-Targeted NADPH Oxidase Mediates Antifungal Host Defense and Regulation of Acute Inflammation in Mice.</a> The Journal of Immunology. 190, 4175-4184 (2013).</li><li>Schaffner, A., Douglas, H., Braude, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Selective+protection+against+conidia+by+mononuclear+and+against+mycelia+by+polymorphonuclear+phagocytes+in+resistance+to+Aspergillus.+Observations+on+these+two+lines+of+defense+in+vivo+and+in+vitro+with+human+and+mouse+phagocytes.">Selective protection against conidia by mononuclear and against mycelia by polymorphonuclear phagocytes in resistance to Aspergillus. Observations on these two lines of defense in vivo and in vitro with human and mouse phagocytes.</a> J Clin Invest. 69, 617-631 (1982).</li><li>Philippe, B., 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=Killing+of+Aspergillus+fumigatus+by+alveolar+macrophages+is+mediated+by+reactive+oxidant+intermediates.">Killing of Aspergillus fumigatus by alveolar macrophages is mediated by reactive oxidant intermediates.</a> Infect Immun. 71, 3034-3042 (2003).</li><li>Cornish, E. J., 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=Reduced+nicotinamide+adenine+dinucleotide+phosphate+oxidase-independent+resistance+to+Aspergillus+fumigatus+in+alveolar+macrophages.">Reduced nicotinamide adenine dinucleotide phosphate oxidase-independent resistance to Aspergillus fumigatus in alveolar macrophages.</a> J Immunol. 180, 6854-6867 (2008).</li><li>Jennings, J. H., Linderman, D. J., Hu, B., Sonstein, J., Curtis, J. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Monocytes+recruited+to+the+lungs+of+mice+during+immune+inflammation+ingest+apoptotic+cells+poorly.">Monocytes recruited to the lungs of mice during immune inflammation ingest apoptotic cells poorly.</a> Am J Respir Cell Mol Biol. 32, 108-117 (2005).</li><li>Davidson, B. 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=DISCRIMINATION+OF+RESIDENT+AND+INFILTRATED+ALVEOLAR+MACROPHAGES+BY+FLOW+CYTOMETRY+IN+INFLUENZA+A+VIRUS-INFECTED+MICE.">DISCRIMINATION OF RESIDENT AND INFILTRATED ALVEOLAR MACROPHAGES BY FLOW CYTOMETRY IN INFLUENZA A VIRUS-INFECTED MICE.</a> Experimental Lung Research. 31, 323-339 (2005).</li><li>Gelderman, K. 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=Macrophages+suppress+T+cell+responses+and+arthritis+development+in+mice+by+producing+reactive+oxygen+species.">Macrophages suppress T cell responses and arthritis development in mice by producing reactive oxygen species.</a> J Clin Invest. 117, 3020-3028 (2007).</li><li>Pizzolla, 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=Reactive+oxygen+species+produced+by+the+NADPH+oxidase+2+complex+in+monocytes+protect+mice+from+bacterial+infections.">Reactive oxygen species produced by the NADPH oxidase 2 complex in monocytes protect mice from bacterial infections.</a> J Immunol. 188, 5003-5011 (2012).</li><li>Luther, K., 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=Characterisation+of+the+phagocytic+uptake+of+Aspergillus+fumigatus+conidia+by+macrophages.">Characterisation of the phagocytic uptake of Aspergillus fumigatus conidia by macrophages.</a> Microbes and Infection. 10, 175-184 (2008).</li><li>Geunes-Boyer, 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=Surfactant+Protein+D+Increases+Phagocytosis+of+Hypocapsular+Cryptococcus+neoformans+by+Murine+Macrophages+and+Enhances+Fungal+Survival.">Surfactant Protein D Increases Phagocytosis of Hypocapsular Cryptococcus neoformans by Murine Macrophages and Enhances Fungal Survival.</a> Infection and Immunity. 77, 2783-2794 (2009).</li><li>Garc&#237;a-Rodas, R., Gonz&#225;lez-Camacho, F., Rodr&#237;guez-Tudela, J. L., Cuenca-Estrella, M., Zaragoza, O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Interaction+between+Candida+krusei+and+Murine+Macrophages+Results+in+Multiple+Outcomes,+Including+Intracellular+Survival+and+Escape+from+Killing.">The Interaction between Candida krusei and Murine Macrophages Results in Multiple Outcomes, Including Intracellular Survival and Escape from Killing.</a> Infection and Immunity. 79, 2136-2144 (2011).</li><li>Ryan, L. K., Vermeulen, M. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Alveolar+macrophages+from+C3H/HeJ+mice+show+sensitivity+to+endotoxin.">Alveolar macrophages from C3H/HeJ mice show sensitivity to endotoxin.</a> Am J Respir Cell Mol Biol. 12, 540-546 (1995).</li><li>Redente, E. F., 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=Differential+polarization+of+alveolar+macrophages+and+bone+marrow-derived+monocytes+following+chemically+and+pathogen-induced+chronic+lung+inflammation.">Differential polarization of alveolar macrophages and bone marrow-derived monocytes following chemically and pathogen-induced chronic lung inflammation.</a> J Leukoc Biol. 88, 159-168 (2010).</li><li>Schaffner, A., Douglas, H., Braude, A. I., Davis, C. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Killing+of+Aspergillus+spores+depends+on+the+anatomical+source+of+the+macrophage.">Killing of Aspergillus spores depends on the anatomical source of the macrophage.</a> Infect Immun. 42, 1109-1115 (1983).</li><li>Goodridge, H. 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=Differential+use+of+CARD9+by+dectin-1+in+macrophages+and+dendritic+cells.">Differential use of CARD9 by dectin-1 in macrophages and dendritic cells.</a> J Immunol. 182, 1146-1154 (2009).</li></ol>

Usando questo approccio ex vivo per analisi dell&#39;attività antifungina di macrofagi insieme in vivo sfida fungina, abbiamo precedentemente dimostrato che NADPH ossidasi in macrofagi svolge un ruolo importante nella difesa contro A. fumigatus 4. L&#39;uso di isolati macrofagi alveolari ci permette di concentrarsi sulla loro attività antifungina in assenza di altre cellule immunitarie (ad esempio, neutrofili reclutati) che difendersi Aspergillus in vivo. <p ...

Macrofagi alveolari sono i fagociti di prima linea che incontrano i microbi inalati. I macrofagi riconoscono motivi Aspergillus dai recettori di riconoscimento patogeni, ingerire e limitano la crescita delle spore per via inalatoria (conidi), e avviare le risposte infiammatorie. Il ossidasi dei fagociti NADPH converte l&#39;ossigeno molecolare per anione superossido e valle intermedi reattivi dell&#39;ossigeno (ROI). Malattia granulomatosa cronica (CGD) è una malattia ereditaria della NADPH ossidasi caratterizzata da infezioni batteriche e fungine gravi e le risposte infiammatorie eccessive. Anche se NADPH ossidasi è fondamentale per neutrofilo-mediato difesa ospite 1-3, l&#39;importanza della NADPH ossidasi nei macrofagi non è ben definito. Precedenti studi hanno dimostrato che i macrofagi alveolari ingerire ed uccidere le spore di Aspergillus, considerando che i neutrofili principalmente di mira il palco ife 5. Tuttavia, ci sono stati Resul contrastantits come il ruolo della NADPH ossidasi in macrofagi nel controllare la crescita di A. fumigatus spore 6, 7. L&#39;obiettivo di questo metodo è stato quello di delineare il ruolo specifico della NADPH ossidasi nei macrofagi nel mediare difesa dell&#39;ospite contro A. spore fumigatus. Abbiamo trovato che NADPH ossidasi in macrofagi alveolari controlla la crescita di fagocitato A. fumigatus spore 4. Per modellare più strettamente la risposta dell&#39;ospite ai funghi per via inalatoria, abbiamo usato macrofagi alveolari raccolte da lavaggio broncoalveolare da topi non stimolati subito dopo il sacrificio. L&#39;uso di isolati macrofagi alveolari ci ha consentito di concentrarsi sulla loro attività antifungina in assenza di altre cellule immunitarie (ad esempio, neutrofili reclutati) che difendersi Aspergillus in vivo. In questo metodo, macrofagi alveolari sono state colorate in vivo prima della raccolta in modo da minimizzare la quantità di manipolazione post-raccolta. &lt;/ P&gt; Un altro vantaggio di questo protocollo è l&#39;uso di un GFP che esprimono A. ceppo fumigatus. Questo fungo esprime GFP dalla fase conidi attraverso la fase di ife e non ha bisogno di ulteriori colorazione. Per ottenere immagini con maggiore dettaglio, abbiamo scelto microscopia confocale che permette la ricostruzione di strutture tridimensionali delle immagini ottenute. Ricostruzione tridimensionale dà la capacità di distinguere tra ife crescono intorno piuttosto che in o attraverso un macrofago. Conidi può anche essere precisamente distinto come intracellulare contro extracellulare.

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		<tr height="63">
			<td height="63" style="height:63px;width:216px;">PKH26 Red Fluorescent Cell Linker Kit for General Cell Membrane Labeling</td>
			<td style="width:71px;">Sigma</td>
			<td style="width:119px;">PKH26GL-1KT</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">2,2,2 Tribromoethanol</td>
			<td style="width:71px;">Sigma</td>
			<td style="width:119px;">T48402</td>
			<td>Used to make AvertinAnesthetic</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">2-methyl-2-butanol</td>
			<td style="width:71px;">Sigma</td>
			<td style="width:119px;">A-1685</td>
			<td>Used to make AvertinAnesthetic</td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:216px;">Dulbecco&#39;s Phosphate Buffered Saline (without calcium and magnesium)</td>
			<td style="width:71px;">Corning</td>
			<td style="width:119px;">21-031-CV</td>
			<td></td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;width:216px;">NH4CL</td>
			<td style="width:71px;">Sigma</td>
			<td style="width:119px;">A0171</td>
			<td>Used to make ACK red cell lysis buffer</td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;width:216px;">KHCO3</td>
			<td style="width:71px;">Sigma</td>
			<td style="width:119px;">P91444</td>
			<td>Used to make ACK red cell lysis buffer</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">EDTA</td>
			<td style="width:71px;">Sigma</td>
			<td style="width:119px;">E6758</td>
			<td>Used to make ACK red cell lysis buffer</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">22 gauge X 1.00 inch i.v. catheter</td>
			<td style="width:71px;">BD&#160;</td>
			<td align="right" style="width:119px;">381523</td>
			<td>Insyte Autoguard Winged</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">insulin syringes</td>
			<td style="width:71px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">6 ml syringes</td>
			<td style="width:71px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">4-way stopcock</td>
			<td style="width:71px;">Fisher Scientific</td>
			<td style="width:119px;">50-700-077</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">suture thread</td>
			<td style="width:71px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">50 ml conical centrifuge tubes</td>
			<td style="width:71px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">gauze sponges (4 inch square)</td>
			<td style="width:71px;"></td>
			<td style="width:119px;"></td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;width:216px;">RPMI 1640 with L-glutamine</td>
			<td style="width:71px;">Corning</td>
			<td style="width:119px;">10-040-CV</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Fetal Bovine Serum</td>
			<td style="width:71px;">Hyclone</td>
			<td style="width:119px;">SH30396.03</td>
			<td>heat inactivated</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Hema 3 Stain Set</td>
			<td style="width:71px;">Fisher Scientific</td>
			<td style="width:119px;">22-122-911</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Cytoseal 60</td>
			<td style="width:71px;">Thermo Scientific</td>
			<td style="width:119px;">8310-4</td>
			<td></td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:216px;">Sabouraud Brain Heart Infusion Agar with Chloramphenicol and Gentamicin Slants</td>
			<td style="width:71px;">BD&#160;</td>
			<td align="right" style="width:119px;">297252</td>
			<td></td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:216px;">Aspergillus fumigatous strain expressing green fluorescent protein</td>
			<td style="width:71px;"></td>
			<td style="width:119px;"></td>
			<td>provided by Dr Margo Moore, Simon Fraser University, Burnaby, BC, Canada</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">100 micron Cell Strainers</td>
			<td style="width:71px;">BD Falcon</td>
			<td style="width:119px;">64753-00</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;width:216px;">scissors</td>
			<td style="width:71px;"></td>
			<td style="width:119px;"></td>
			<td></td>
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			<td height="21" style="height:21px;width:216px;">forceps</td>
			<td style="width:71px;"></td>
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			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Biological Safety Cabinet Class II</td>
			<td style="width:71px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Sorval ST40 Centrifuge with swing bucket rotor</td>
			<td style="width:71px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Leica DM1000 light microscope</td>
			<td style="width:71px;"></td>
			<td style="width:119px;"></td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Hemocytometer</td>
			<td style="width:71px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Cytospin 2 centrifuge</td>
			<td style="width:71px;">Thermo Scientific</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;width:216px;">Cell Culture Incubator</td>
			<td style="width:71px;"></td>
			<td style="width:119px;"></td>
			<td>37 &#176;C, 5%&#160; CO2</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">22X22 mm micro cover glass</td>
			<td style="width:71px;">VWR</td>
			<td style="width:119px;">48366-227</td>
			<td>glass coverslips</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">6 well Cell Culture Plates</td>
			<td style="width:71px;">Corning</td>
			<td align="right" style="width:119px;">3506</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">10% Neutral Buffered Formalin</td>
			<td style="width:71px;">VWR</td>
			<td style="width:119px;">BDH0502-1LP</td>
			<td></td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:216px;">Vectashield Mounting medium with DAPI</td>
			<td style="width:71px;">Vector Laboratories</td>
			<td style="width:119px;">H-1200</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Leica TCS SP2 system with a laser point scanner</td>
			<td style="width:71px;"></td>
			<td style="width:119px;"></td>
			<td>Mounted on DMIRE2 fluorescence microscope</td>
		</tr>
	</tbody>
</table>

assessing anti-fungal activity of isolated alveolar macrophages by confocal microscopy

Il polmone è un&#39;interfaccia in cui cellule ospiti sono regolarmente esposti ai microbi e prodotti microbici. Macrofagi alveolari sono i fagociti di prima linea che incontrano i funghi per via inalatoria e altri microbi. Macrofagi e altre cellule immunitarie riconoscono motivi Aspergillus dai recettori di riconoscimento patogeno e avviare risposte infiammatorie a valle. Il ossidasi dei fagociti NADPH genera intermedi reattivi dell&#39;ossigeno (ROI) ed è fondamentale per la difesa ospite. Anche se NADPH ossidasi è fondamentale per neutrofilo-mediato difesa ospite 1-3, l&#39;importanza della NADPH ossidasi nei macrofagi non è ben definito. L&#39;obiettivo di questo studio è stato quello di delineare il ruolo specifico della NADPH ossidasi nei macrofagi nel mediare difesa dell&#39;ospite contro A. fumigatus. Abbiamo trovato che NADPH ossidasi in macrofagi alveolari controlla la crescita di fagocitato A. fumigatus spore 4. Qui, descriviamo un metodo per valutare la capacità di un topomacrofagi lveolar (AMS) per controllare la crescita delle spore di Aspergillus fagocitati (conidi). Macrofagi alveolari sono macchiati in vivo e dieci giorni più tardi isolati da topi da lavaggio broncoalveolare (BAL). I macrofagi sono placcati su vetrini, poi seminate con la proteina fluorescente verde (GFP) che esprimono A. spore fumigatus. A volte specificato, le cellule vengono fissate e il numero di macrofagi intatte con spore fagocitati è valutata mediante microscopia confocale.

<html> Raccolta dei macrofagi alveolari da BAL da topi non stimolati si tradurrà in una popolazione pura&gt; 95% in base citologia. Il 3 e 7 ore (Figura 2) punti temporali precedono la fase di ife di crescita fungina e consentono un confronto di efficienza fagocitaria dei conidi tra i genotipi. Il punto di tempo 14 hr è dove genotipi possono essere confrontati per quanto riguarda la capacità di inibire il passaggio di conidi fagocitato alla fase hyphal tessuto invasivo (Figura 3). Mac...

Watch this Scientific Journal Video about Assessing Anti-fungal Activity of Isolated Alveolar Macrophages by Confocal Microscopy at JoVE.com

Assessing Anti-fungal Activity of Isolated Alveolar Macrophages by Confocal Microscopy

Valutare l&#39;attività anti-fungina isolato Alveolar macrofagi dal Microscopia confocale

Un metodo per valutare la capacità di isolato macrofagi alveolari del mouse per controllare la crescita di spore di Aspergillus fagocitati mediante microscopia confocale.

The lung is an interface where host cells are routinely exposed to microbes and microbial products. Alveolar macrophages are the first-line phagocytic ...

Research

JoVE Journal

Immunology and Infection

Microwave Assisted Rapid Diagnosis of Plant Virus Diseases by Transmission Electron Microscopy

Forno a microonde Assist diagnosi rapida di malattie virali piante da microscopia elettronica a trasmissione

Investigations of ultrastructural changes induced by viruses are often necessary to clearly identify viral diseases in plants. With conventional sample ...

Ricerca

Immunologia e infezioni

Intravital Microscopy of the Spleen: Quantitative Analysis of Parasite Mobility and Blood Flow

Microscopia intravitale della Milza: analisi quantitativa della mobilità Parasite e del flusso sanguigno

The advent of intravital microscopy in experimental rodent malaria models has allowed major advances to the knowledge of parasite-host interactions 1,2. ...

Real-time Imaging of Myeloid Cells Dynamics in ApcMin/+ Intestinal Tumors by Spinning Disk Confocal Microscopy

Real-time Imaging of Myeloid Cells Dynamics in ApcMin/+ Intestinal Tumors by Spinning Disk Confocal Microscopy

In tempo reale Imaging di cellule mieloidi Dynamics in<em&gt; Apc<sup&gt; Min / +</sup</em&gt; Intestinale tumori da Spinning Disk Microscopia confocale

Myeloid cells are the most abundant immune cells within tumors and have been shown to promote tumor progression. Modern intravital imaging techniques ...

Proteomic Profiling of Macrophages by 2D Electrophoresis

Proteomica Profiling dei macrofagi da 2D elettroforesi

The goal of the two-dimensional (2D) electrophoresis protocol described here is to show how to analyse the phenotype of human cultured macrophages. The ...

Generation and Identification of GM-CSF Derived Alveolar-like Macrophages and Dendritic Cells From Mouse Bone Marrow

Generazione e identificazione di GM-CSF Derivato alveolo-come macrofagi e dendritiche cellule da midollo osseo di topo

Macrophages and dendritic cells (DCs) are innate immune cells found in tissues and lymphoid organs that play a key role in the defense against pathogens. ...

Phenotypic Characterization of Macrophages from Rat Kidney by Flow Cytometry

Caratterizzazione fenotipica dei macrofagi dal rene di ratto mediante citometria di flusso

There is increasing evidence suggesting the important role of inflammation and, subsequently, macrophages in the development and progression of renal ...

Isolation, Characterization, and Purification of Macrophages from Tissues Affected by Obesity-related Inflammation

Isolamento, caratterizzazione e purificazione dei macrofagi dai tessuti colpiti da infiammazione obesità

Obesity promotes a chronic inflammatory state that is largely mediated by tissue-resident macrophages as well as monocyte-derived macrophages. ...

Visualizing Macrophage Extracellular Traps Using Confocal Microscopy

Visualizzazione del macrofago trappole extracellulari mediante microscopia confocale

A primary method used to define the presence of neutrophil extracellular traps (NETs) is confocal microscopy. We have modified established confocal ...

Isolation and In Vitro Culture of Murine and Human Alveolar Macrophages

Isolation and In Vitro Culture of Murine and Human Alveolar Macrophages

Isolamento e coltura In Vitro dei macrofagi alveolari murini ed umani

Alveolar macrophages are terminally differentiated, lung-resident macrophages of prenatal origin. Alveolar macrophages are unique in their long life and ...

Time-lapse 3D Imaging of Phagocytosis by Mouse Macrophages

Time-lapse Imaging 3D della fagocitosi dai macrofagi del topo

Phagocytosis plays a key role in host defense, as well as in tissue development and maintenance, and involves rapid, receptor-mediated rearrangements of ...