Name: Tomografia al microscopio elettronico a scansione a trasmissione in virologia: imaging 3D di campioni congelati ad alta pressione, liofilizzati (Video) | JoVE Journal
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Description: Impara la tomografia STEM in virologia per imaging 3D di cellule infette. Questo metodo combina congelamento ad alta pressione e sostituzione criogenica per visualizzare alterazioni ultrastrutturali con maggiore dettaglio.

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</ol>

<table><tbody><tr><td>1-Esadeceno</td><td>Merck KGaA</td><td>8220640500</td><td></td></tr><tr><td>Griglia di rame da 200 con barre parallele</td><td>Plano GmbH</td><td>G2014C</td><td></td></tr><tr><td>37 & deg; Camera di asciugatura a C</td><td>Binder GmbH</td><td>9010-0323</td><td></td></tr><tr><td>Acetone</td><td>VWR international GmbH</td><td>20066330</td><td></td></tr><tr><td>DDSA</td><td>Scienze della microscopia elettronica</td><td>13710</td><td></td></tr><tr><td>Coltello a diamante</td><td>Diatome Ltd</td><td>https://www.diatomeknives.com/ultra-35</td><td></td></tr><tr><td>DMP-30</td><td>Scienze della microscopia elettronica</td><td>13600</td><td></td></tr><tr><td>Embed-812 Resina</td><td>Scienze della microscopia elettronica</td><td>14900</td><td></td></tr><tr><td>Strumenti EM</td><td>TVIPS GmbH</td><td>Software TVIPS: EM-Tools; https://www.tvips.com/imaging-software/em-tools/</td><td></td></tr><tr><td>Etanolo al 100%</td><td>Merck KGaA</td><td>1.00983.2511</td><td></td></tr><tr><td>Carta filtro</td><td>VWR international GmbH</td><td>516-0812</td><td></td></tr><tr><td>Flaschen mit Schnappdeckelglas 11 mL</td><td>VWR international GmbH</td><td>548-0625</td><td></td></tr><tr><td>Formvar</td><td>Scienze della microscopia elettronica</td><td>15830-25</td><td></td></tr><tr><td>Unità di sostituzione del congelamento Leica EM AFS2</td><td>Leica Microsystems GmbH</td><td>https://www.leica-microsystems.com/products/sample-preparation-for-electron-microscopy/p/leica-em-afs2/</td><td></td></tr><tr><td>Glod Sol 25 nm</td><td>Reagenti e Reagenti d'Oro AURION Immuno Accessori</td><td>425.011</td><td></td></tr><tr><td>Buca di pino dorata e oslash; 2000 e MU; m</td><td>Science Services GmbH</td><td>GA2000-AU</td><td></td></tr><tr><td>Graiticules Buchstaben-Netzchen Kupfer</td><td>Plano GmbH</td><td>NH6C</td><td></td></tr><tr><td>Piastra riscaldante</td><td>MEDAX GmbH & Co. KG</td><td>12501</td><td></td></tr><tr><td>Sistema di vaporizzazione ad alto vuoto BAF 300</td><td>BAL-TEC AG</td><td></td><td></td></tr><tr><td>Congelatore ad alta pressione Wohlwend HPF Compact 01</td><td>Ufficio di Ingegneria M. Wohlwend GmbH</td><td>https://www.wohlwend-hpf.ch/index.html</td><td></td></tr><tr><td>Serbatoio di azoto liquido</td><td>H.Erben GmbH</td><td>94500</td><td></td></tr><tr><td>Azoto (fluido)</td><td>VWR international GmbH</td><td>-</td><td></td></tr><tr><td>NMA</td><td>Scienze della microscopia elettronica</td><td>19000</td><td></td></tr><tr><td>Tetrossido di osmio</td><td>ChemPur Feinchemikalien und Forschungsbedarf GmbH</td><td>006051</td><td></td></tr><tr><td>Forno a 120 °C; C</td><td>Binder GmbH</td><td>9010-0194</td><td></td></tr><tr><td>PELCO easi-Glow</td><td>Ted Pella, Inc.</td><td>91000</td><td></td></tr><tr><td>Poly-L-Lysin 0,1% rispetto all'aq. Soluzione</td><td>Ted Pella, Inc.</td><td>18026</td><td></td></tr><tr><td>Tubi Safe-Lock 0,5 mL</td><td>Eppendorf SE</td><td>0030 121.023</td><td></td></tr><tr><td>Contenitori campione (capsule Beem)</td><td>Plano GmbH</td><td>G360-1</td><td></td></tr><tr><td>Disco zaffiro diametro 3 x 0,16 mm</td><td>Ufficio di Ingegneria M. Wohlwend GmbH</td><td>500</td><td></td></tr><tr><td>Ultramicrotomo Leica EM UC7</td><td>Leica Microsystems GmbH</td><td>https://www.leica-microsystems.com/de/produkte/em-probenvorbereitung/p/leica-em-uc7/</td><td></td></tr><tr><td>Acetato di uranyle</td><td>Riedel de Ha&euml; n AG</td><td>31697</td><td></td></tr><tr><td>Camera a luce UV</td><td>Dinies</td><td>ELG100S</td><td></td></tr></tbody></table>

tomografia al microscopio elettronico a scansione a trasmissione in virologia: imaging 3d di campioni congelati ad alta pressione, liofilizzati

Scanning Transmission Electron Microscopy Tomography in Virology: 3D Imaging of High-pressure Frozen, Freeze-substituted Samples

Tomografia al microscopio elettronico a scansione a trasmissione in virologia: imaging 3D di campioni congelati ad alta pressione, liofilizzati

Electron microscopy (EM) and especially three-dimensional (3D) EM techniques have become established methods in structural virology. Investigation of ...

scanning-transmission-electron-microscopy-tomography-virology-3d

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Questo rapporto descrive un protocollo di preparazione del campione e condizioni di imaging specifiche per eseguire la tomografia elettronica a scansione di trasmissione di campioni biologici spessi.

Preparazione e di osservazione di campioni biologici spessi da Scanning Transmission Electron Tomografia

Questo protocollo viene descritto come visualizzare la compattazione DNA transitoria nei cianobatteri. Coltivazione sincrono, monitoraggio di microscopia di fluorescenza, congelamento rapido e alta tomografia di cryo-elettrone di tensione sono usati. Un protocollo per queste metodologie è presentato, e sono discussi gli sviluppi e le applicazioni future.

Visualizzazione di compattazione del DNA nei cianobatteri tramite tomografia ad alta tensione Cryo-elettrone

Qui viene presentato un protocollo che descrive le fasi di preparazione del campione e di raccolta dei dati richieste nella tomografia a raggi X criomoscido morbido (SXT) per visualizzare l'ultrastruttura di intere cellule crioconservate con una risoluzione di 25 nm a mezzo passo.

Una descrizione cartografica 3D della cellula mediante tomografia a raggi X Cryo Soft

Presentiamo un protocollo per la preparazione lamellare di campioni biologici surgelati a tuffo mediante microlavorazione a fascio ionico crio-focalizzato per studi strutturali ad alta risoluzione di macromolecole in situ con tomografia crioeletnico. Il protocollo presentato fornisce linee guida per la preparazione di lamelle di alta qualità ad alta riproducibilità per la caratterizzazione strutturale di macromolecole all'interno del Saccharomyces cerevisiae.

Preparazione e microlavorazione Cryo-FIB di Saccharomyces cerevisiae per Tomografia Crioeletnico

Qui vengono descritti i protocolli per preparare assemblaggi di virus adatti per l'analisi liquido-EM e crio-EM su scala nanometrica utilizzando la microscopia elettronica a trasmissione.

Miglioramento dell'imaging ad alta risoluzione di assemblaggi di virus in liquidi e ghiaccio

La tomografia crio-STEM fornisce un mezzo per visualizzare organelli di cellule intatte senza incorporamento, sezionamento o altre preparazioni invasive. La risoluzione 3D ottenuta è attualmente nell'intervallo di pochi nanometri, con un campo visivo di diversi micrometri e uno spessore accessibile nell'ordine di 1 μm.

Visualizzazione di organelli in situ mediante tomografia crio-STEM

Impara la tomografia STEM in virologia per imaging 3D di cellule infette. Questo metodo combina congelamento ad alta pressione e sostituzione criogenica per visualizzare alterazioni ultrastrutturali con maggiore dettaglio.

Tomografia al microscopio elettronico a scansione a trasmissione in virologia: imaging 3D di campioni congelati ad alta pressione, liofilizzati (Video) | JoVE Journal

Tomografia al microscopio elettronico a scansione a trasmissione in virologia: imaging 3D di campioni congelati ad alta pressione, liofilizzati

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