{"id":1136,"date":"2022-01-18T16:04:58","date_gmt":"2022-01-18T15:04:58","guid":{"rendered":"https:\/\/michelecrevecoeur.ch\/?page_id=1136"},"modified":"2022-02-27T18:25:41","modified_gmt":"2022-02-27T17:25:41","slug":"scanning-electron-microscopy","status":"publish","type":"page","link":"https:\/\/michelecrevecoeur.ch\/?page_id=1136","title":{"rendered":"Scanning Electron Microscopy"},"content":{"rendered":"\t\t
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Scanning Electron Microscope<\/span><\/span><\/p>

In the Scanning Electron Microscope, a beam of high \u2013 energy electrons is emitted from an electron gun and accelerated down to an energy up to 30 keV. It passes through a combination of magnetic lenses and apertures that focus the electrons into a small probe. This rapidly scans the surface of the sample. The interactions between electrons and the sample surface produces various signals detected by appropriate detectors which allow to obtain information on the surface topography but also on compositional features of the sample. The resolution of modern SEM is about 10 nm, varying from 1 to 20 nm, and magnifications of 200.000x up to 1.000.000 x can be achieved. The SEM permits non-destructive examen of the specimen.
As for TEM a vacuum is required in a SEM, to protect filament and to avoid collision between electrons and gas molecules. To improve quality of images the surface of specimen is coated with a thin layer of metal, generally gold. The penetration of electrons in the matter is very low, particularly after coating.
Some biological samples relatively dry can be observed directly without metallization: seeds, pollen grains. On the contrary soft and hydrated samples need a particular preparation including chemical fixation, washing, dehydration, drying and finally metallization.<\/p>

\u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 Exemple of protocol of specimen preparation\u00a0<\/span><\/span>
<\/span>
<\/span>All the steps of the protocol take place at laboratory temperature.<\/p>

\u00a0\u25cf 1st\u00a0fixation:\u00a0 2.5% glutaraldehyde in sodium cacodylate buffer (0.1M; pH 7.0) for 2h
\u00a0\u25cf Washing in buffer cacodylate 4 x 15 min
\u00a0\u25cf 2nd\u00a0fixation : 1% OsO4 in cacodylate buffer for 2 h
\u00a0\u25cf\u00a0Washing in Cacodylate buffer\u00a0 2 x 15 min then with milliQ\u00a0water 2 x 15 min
\u00a0\u25cf\u00a0Dehydration in ethanol :\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0<\/p>

\u00a0 \u00a0 \u00a0 \u00a0 \u00a0\u00a0 Ethanol 30 \u00a0\u00a0 \u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0 20 min\u00a0\u00a0
\u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0Ethanol 50 \u00a0\u00a0 \u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0 20 min\u00a0\u00a0
\u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0Ethanol 70 \u00a0\u00a0 \u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0 20 min
\u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0Ethanol 90 \u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0 20 min\u00a0 \u00a0 \u00a0\u00a0
\u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0Ethanol 100 I\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0 3 x 20 min\u00a0 \u00a0<\/p>

\u00a0\u25cf\u00a0Critical air drying
\u00a0\u25cf\u00a0Metallization\u00a0: coating with a thin layer of gold using a \u00ab\u00a0JEOL 1200 – Fine coater<\/a>\u00a0\u00bb from the Bioimaging Platform, Sciences Faculty, Sciences II, University of Geneva.
\u00a0\u25cf\u00a0Observations: Jeol JSM-6510 LV microscope \u2013\u00a0see the Protocol for using \u2013\u00a0written in collaboration with St\u00e9phane Hagmann (research auxiliary from May to August 2011).<\/p>

Plant organs imaged with the Scanning Electron Microscope\u00a0<\/span><\/span><\/span>JEOL JSM 6510LV<\/span><\/span><\/span><\/a><\/p>

Imaging<\/span>\u00a0<\/span>:\u00a0St\u00e9phane Hagmann (research auxiliary; Kilian Anderegg (apprentice); Mich\u00e8le Cr\u00e8vecoeur (Lecturer; Microscopist)
Annotations & picture’s comments<\/span><\/span>\u00a0: Mich\u00e8le Cr\u00e8vecoeur<\/p><\/div>

\u00a0<\/div><\/div>

Leaves<\/span>\u00a0:
\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0Arabidopsis thaliana<\/span><\/em><\/a>
\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0Origanum vulgare<\/span><\/em><\/a>
\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0Rosmarinus officinalis<\/em><\/span><\/a>
\u00a0 \u00a0 \u00a0 \u00a0\u00a0Salvia officinalis<\/em><\/span><\/a>\u00a0\u00a0
\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0Triticum durum
<\/em><\/span><\/a>\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0Drosera linearis<\/em><\/span><\/a><\/p><\/div>

Stem :
\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0S<\/span><\/em>alvia officinalis<\/span><\/em><\/a>
<\/span><\/em><\/a><\/span>Roots<\/span>:
\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0<\/span>Zea mays<\/span><\/em><\/a>
<\/em><\/a>Embryo:
\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0Arabidopsis thaliana<\/em><\/span><\/a><\/p><\/div>

Flower:<\/span>
\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0Arabidopsis<\/em><\/a><\/span><\/p>

\u00a0<\/div><\/div>
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\u00a0Scanning Electrons Microscopes in Geneva<\/strong><\/span><\/p>

Dr Andr\u00e9 Piuz<\/span>
<\/strong><\/a>Geneva’s Natural History Museum<\/span>
<\/span>Scanning Electrons Microscopes’ laboratories and X-Ray microanalysis<\/span><\/p>

Bioimaging Center<\/a>
Faculty of Sciences (<\/span>Sciences II) – Geneva University<\/span><\/p>

Prof. Rossana Martini<\/a>
Earth Science<\/p>\t\t\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<\/div>\n\t\t","protected":false},"excerpt":{"rendered":"

Scanning Electron Microscope In the Scanning Electron Microscope, a beam of high \u2013 energy electrons is emitted from an electron gun and accelerated down to an energy up to 30 keV. It passes through a combination of magnetic lenses and apertures that focus the electrons into a small probe. This rapidly scans the surface of […]<\/p>\n","protected":false},"author":1,"featured_media":0,"parent":0,"menu_order":0,"comment_status":"closed","ping_status":"closed","template":"","meta":{"site-sidebar-layout":"no-sidebar","site-content-layout":"page-builder","ast-site-content-layout":"default","site-content-style":"default","site-sidebar-style":"default","ast-global-header-display":"","ast-banner-title-visibility":"","ast-main-header-display":"","ast-hfb-above-header-display":"","ast-hfb-below-header-display":"","ast-hfb-mobile-header-display":"","site-post-title":"disabled","ast-breadcrumbs-content":"","ast-featured-img":"disabled","footer-sml-layout":"","ast-disable-related-posts":"","theme-transparent-header-meta":"","adv-header-id-meta":"","stick-header-meta":"","header-above-stick-meta":"","header-main-stick-meta":"","header-below-stick-meta":"","astra-migrate-meta-layouts":"default","ast-page-background-enabled":"default","ast-page-background-meta":{"desktop":{"background-color":"var(--ast-global-color-4)","background-image":"","background-repeat":"repeat","background-position":"center center","background-size":"auto","background-attachment":"scroll","background-type":"","background-media":"","overlay-type":"","overlay-color":"","overlay-opacity":"","overlay-gradient":""},"tablet":{"background-color":"","background-image":"","background-repeat":"repeat","background-position":"center center","background-size":"auto","background-attachment":"scroll","background-type":"","background-media":"","overlay-type":"","overlay-color":"","overlay-opacity":"","overlay-gradient":""},"mobile":{"background-color":"","background-image":"","background-repeat":"repeat","background-position":"center center","background-size":"auto","background-attachment":"scroll","background-type":"","background-media":"","overlay-type":"","overlay-color":"","overlay-opacity":"","overlay-gradient":""}},"ast-content-background-meta":{"desktop":{"background-color":"var(--ast-global-color-5)","background-image":"","background-repeat":"repeat","background-position":"center center","background-size":"auto","background-attachment":"scroll","background-type":"","background-media":"","overlay-type":"","overlay-color":"","overlay-opacity":"","overlay-gradient":""},"tablet":{"background-color":"var(--ast-global-color-5)","background-image":"","background-repeat":"repeat","background-position":"center center","background-size":"auto","background-attachment":"scroll","background-type":"","background-media":"","overlay-type":"","overlay-color":"","overlay-opacity":"","overlay-gradient":""},"mobile":{"background-color":"var(--ast-global-color-5)","background-image":"","background-repeat":"repeat","background-position":"center center","background-size":"auto","background-attachment":"scroll","background-type":"","background-media":"","overlay-type":"","overlay-color":"","overlay-opacity":"","overlay-gradient":""}},"footnotes":""},"class_list":["post-1136","page","type-page","status-publish","hentry"],"yoast_head":"\nScanning Electron Microscopy - Mich\u00e8le Cr\u00e8vecoeur<\/title>\n<meta name=\"robots\" content=\"index, follow, max-snippet:-1, max-image-preview:large, max-video-preview:-1\" \/>\n<link rel=\"canonical\" href=\"https:\/\/michelecrevecoeur.ch\/?page_id=1136\" \/>\n<meta property=\"og:locale\" content=\"fr_FR\" \/>\n<meta property=\"og:type\" content=\"article\" \/>\n<meta property=\"og:title\" content=\"Scanning Electron Microscopy - Mich\u00e8le Cr\u00e8vecoeur\" \/>\n<meta property=\"og:description\" content=\"Scanning Electron Microscope In the Scanning Electron Microscope, a beam of high \u2013 energy electrons is emitted from an electron gun and accelerated down to an energy up to 30 keV. 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