{"id":6727,"date":"2023-05-12T05:48:01","date_gmt":"2023-05-12T05:48:01","guid":{"rendered":"https:\/\/er-c.org\/?page_id=6727"},"modified":"2023-05-31T09:41:22","modified_gmt":"2023-05-31T09:41:22","slug":"sample-visualization","status":"publish","type":"page","link":"https:\/\/er-c.org\/index.php\/facilities-2\/life-science\/cryo-electron-microscopy-facility\/cryo-em-applications\/sample-visualization\/","title":{"rendered":"Sample Visualization"},"content":{"rendered":"\n
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\n\t\t\t\t\t\tSample Visualization\t\t\t\t\t\t<\/h1>\n\t\t\t\t\t\t<\/div>\n<\/div>\n\n\n
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The aim of the initial visualization is to check purity and homogeneity of the sample and it further provides information about sample morphology and dimensions.<\/p>\n\n\n

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Negatively stained samples<\/h3>\n
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Purified proteins, filaments, viruses and other large assemblies can be rapidly visualized and characterized by negative stain electron microscopy using e.g. our Talos 120 microscope<\/a>. Samples are often initially imaged using negative staining EM for optimizing cryogenic single-particle and tomography specimens<\/p>\n<\/div>\n<\/div><\/div><\/div>

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Micrograph of tobacco mosaic virus (TMV) particles as viewed at 92 kX nominal magnification in our room temperature Talos 120 kV.<\/figcaption><\/figure><\/div><\/div><\/div><\/div>

Vitrified cryo-samples<\/h3>\n
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We offer the visualization of vitrified cryo-samples, e.g. protein complex and lipid mixtures for sample characterization. This visualization can also be part of the screening step to assess the sample feasibility for the single-particle data acquisition<\/a> on cryo-EM<\/a>. The plunge-frozen sample can be shipped to the ER-C or be vitrified on site using one of our sample preparation instruments<\/a> like VitrobotTM<\/sup>, VitroJetTM<\/sup>, Manual or LeicaTM<\/sup> Plunge freezer. A small data set can be collected for initial sample characterization that may shed light on the suitability for high-resolution analysis.<\/p>\n<\/div>\n<\/div><\/div><\/div>

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Micrograph of vitrified tobacco mosaic virus (TMV) particles as viewed at 100 kX nominal magnification in our Talos Arctica 200 kV operated at cryo-conditions.<\/figcaption><\/figure><\/div><\/div><\/div><\/div>
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Samples often need multiple rounds of initial screening until requirements for high-resolution cryo-EM are met. These optimization includes adjustments of both, sample conditions (pH, buffer, additives) and blotting conditions (grid type, blotting time, concentration, vitrification procedure).<\/p>\n<\/div>\n<\/div><\/div><\/div><\/div><\/div><\/div>\n\n\n

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The aim of the initial visualization is to check purity and homogeneity of the sample as well as information about sample morphology and dimensions.<\/p>\n","protected":false},"author":2,"featured_media":6737,"parent":6211,"menu_order":0,"comment_status":"closed","ping_status":"closed","template":"page-templates\/pagebuilder.php","meta":{"_themeisle_gutenberg_block_has_review":false,"footnotes":""},"class_list":["post-6727","page","type-page","status-publish","has-post-thumbnail","hentry"],"_links":{"self":[{"href":"https:\/\/er-c.org\/index.php\/wp-json\/wp\/v2\/pages\/6727","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/er-c.org\/index.php\/wp-json\/wp\/v2\/pages"}],"about":[{"href":"https:\/\/er-c.org\/index.php\/wp-json\/wp\/v2\/types\/page"}],"author":[{"embeddable":true,"href":"https:\/\/er-c.org\/index.php\/wp-json\/wp\/v2\/users\/2"}],"replies":[{"embeddable":true,"href":"https:\/\/er-c.org\/index.php\/wp-json\/wp\/v2\/comments?post=6727"}],"version-history":[{"count":24,"href":"https:\/\/er-c.org\/index.php\/wp-json\/wp\/v2\/pages\/6727\/revisions"}],"predecessor-version":[{"id":7052,"href":"https:\/\/er-c.org\/index.php\/wp-json\/wp\/v2\/pages\/6727\/revisions\/7052"}],"up":[{"embeddable":true,"href":"https:\/\/er-c.org\/index.php\/wp-json\/wp\/v2\/pages\/6211"}],"wp:featuredmedia":[{"embeddable":true,"href":"https:\/\/er-c.org\/index.php\/wp-json\/wp\/v2\/media\/6737"}],"wp:attachment":[{"href":"https:\/\/er-c.org\/index.php\/wp-json\/wp\/v2\/media?parent=6727"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}