{"id":610,"date":"2021-10-18T07:00:25","date_gmt":"2021-10-18T07:00:25","guid":{"rendered":"https:\/\/admin.acceleratingscience.com\/atomic-resolution\/2021\/10\/18\/tundra-cryo-tem-questions-2\/"},"modified":"2022-04-14T13:40:22","modified_gmt":"2022-04-14T13:40:22","slug":"tundra-cryo-tem-questions","status":"publish","type":"post","link":"https:\/\/www.thermofisher.com\/blog\/atomic-resolution\/tundra-cryo-tem-questions\/","title":{"rendered":"Answers to the Most Common Questions About Our Tundra Cryo-TEM"},"content":{"rendered":"

At the dawn of 2021, we introduced a new, more accessible solution for cryo-electron microscopy: the Thermo Scientific Tundra Cryo-TEM<\/a>.<\/p>\n

\"Scientist

Jiri Novacek, PhD, Head of Core Facility, CEITEC using a new Tundra Cryo-TEM at Thermo Fisher Scientific in Brno<\/p><\/div>\n

This smaller yet mighty cryo-EM instrument was designed to bring single particle analysis to every biochemistry laboratory<\/a>. But what does that mean?<\/p>\n

You asked. We answered.<\/p>\n

Tundra Cryo-TEM: Commonly Asked Questions and Answers<\/strong><\/h2>\n

What type of user and facility is the Tundra Cryo-TEM most suitable for?<\/em><\/strong><\/h2>\n

<\/h2>\n

The Tundra Cryo-TEM is an excellent choice for new users who are just getting started in cryo-EM.<\/a> This is due to the guided workflow features and easy sample handling functions. It is also a useful addition for experienced users who are looking for an economical and flexible way to expand their cryo-EM sample preparation optimization capabilities.<\/p>\n

Finally, even traditional room temperature microscopists may consider the Tundra Cryo-TEM to diversify their portfolio of services.<\/p>\n

\"Overcoming<\/p>\n

What are the dimensions of the Tundra Cryo-TEM?<\/em><\/strong><\/h2>\n

Unlike other cryo-EM instruments, the\u00a0Tundra Cryo-TEM is built to fit in most standard sized labs without any extensive modifications.<\/p>\n

The entire system can fit into a 4 m X 4.2 m room with a ceiling height of at least 2.74 m.<\/p>\n

This includes the microscope, support cabinets, sample loading station, HT tank and workstation.<\/p>\n

\"Overcoming<\/p>\n

What makes the Tundra Cryo-TEM suitable as a data collection tool, and not just a screening tool?\u00a0\u00a0<\/em><\/strong><\/h2>\n

The Tundra Cryo-TEM is designed to deliver 3D reconstructions at biological relevant resolutions, particularly using\u00a0the\u00a0single particle\u00a0method.<\/p>\n

We have demonstrated\u00a0this through a\u00a0handful\u00a0of examples,\u00a0Gabaa\u00a0receptor at 3.4A, T20S proteasome at\u00a03.0A,\u00a0Adeno-associated Virus\u00a06 at 3.0A, and apoferritin as benchmark protein at 2.6A<\/a>.<\/p>\n

These amazing results are possible because the Tundra Cryo-TEM:<\/p>\n

    \n
  1. Optimizes the optics for high resolution at 100kV.<\/li>\n
  2. Has a\u00a0CMOS detector.\u00a0Not only\u00a0does it have\u00a0enhanced sensitivity and thus\u00a0good\u00a0signal-to-noise ratio at 100kV\u00a0making it\u00a0suited for\u00a0extreme\u00a0dose-sensitive samples in life science,\u00a0but\u00a0also\u00a0it\u00a0saves\u00a0dose fractionations.\u00a0Dose fractionations\u00a0enable\u00a0the teasing out of\u00a0high-resolution\u00a0information in the\u00a0images that are otherwise blurred out by motion and radiation damage.<\/li>\n
  3. Allows the sample in the column to be cooled by\u00a0an extended\u00a0liquid nitrogen Dewar that lasts 72-hours. This enables\u00a0long data collection.<\/li>\n<\/ol>\n

    \"Overcoming<\/p>\n

    Download our infographic – Overcoming Misperceptions About Cryo-EM >><\/a><\/em><\/strong><\/p>\n

    What kind of camera is best for the Tundra Cryo-TEM and 100 kV applications?\u00a0\u00a0\u00a0<\/em><\/strong><\/h2>\n

    Our Tundra Cryo-TEM is equipped with a scintillator-based CMOS camera, Ceta-F. Ceta-F has two major benefits, which makes it the best detector in the market for 100kV applications.<\/p>\n

      \n
    1. Ceta-F has\u00a0a\u00a0very\u00a0good\u00a0Detective\u00a0Quantum Efficiency (DQE) at 100kV.\u00a0Traditionally scintillator-based cameras suffer from poor signal\u00a0when detecting\u00a0incoming electrons with high\u00a0accelerating voltage, like 200kV\u00a0or 300kV, because\u00a0these electrons with high energy\u00a0don\u2019t interact with the scintillator\u00a0too much.\u00a0However,\u00a0electrons at 100kV generate 2x more\u00a0signal\u00a0compared to\u00a0200kV.<\/li>\n
    2. Ceta-F is radiation hard at 100kV.\u00a0Direct\u00a0Electron\u00a0Detectors\u00a0(DED)\u00a0are\u00a0superb\u00a0in\u00a0generating signal, but at the same time\u00a0suffer\u00a0from\u00a0radiation damage. The damage is more severe with\u00a0electrons\u00a0at 100kV. For the similar reasons\u00a0mentioned above, electrons at 100kV interact\u00a0with the sensor more and\u00a0causes\u00a0more damage.\u00a0None of the commonly known DEDs,\u00a0such as\u00a0Thermo Scientific Falcon,\u00a0Gatan\u00a0K3 and DE-20,\u00a0have proven radiation hardness at 100kV.<\/li>\n<\/ol>\n

      What is the throughput for data collection on Tundra Cryo-TEM?\u00a0\u00a0<\/em><\/strong><\/h2>\n

      The throughput is 250-300 images\/hour. The data sets we collected vary from 1,500 images (apoferritin, negative stain, or some other short screening sessions), to 5,000 images (proteins with symmetry), to 10,000 images (asymmetric protein).<\/p>\n

      Can we do multiple shots per hole on Tundra Cryo-TEM?\u00a0<\/em><\/strong><\/h2>\n

      The Tundra Cryo-TEM has two condenser\u00a0lenses\u00a0and a mini-condenser lens. Having two condenser lenses\u00a0means\u00a0the illumination area is proportional to the\u00a0C2 aperture.<\/p>\n

      We\u00a0often use 50um and 70um C2 on the Tundra Cryo-TEM, which results in a 1.25um and 1.75um illumination area. With\u00a0grid type R1.2\/1.3, only one shot is possible on the Tundra Cryo-TEM. With grid type\u00a0R2\/2, when collecting close to the edge of the\u00a0foil hole, it is possible to have multiple shots per hole.\u00a0We have done two shots per hole with 5the\u00a00um C2 aperture\u00a0on R2\/1\u00a0grid.<\/p>\n

      Can we use gold grids for data collection?\u00a0<\/em><\/strong><\/h2>\n

      Yes,\u00a0we can collect data with gold grids on the Tundra Cryo-TEM.<\/p>\n

      However,\u00a0the challenge is with\u00a0the\u00a0automated coma-free alignment routine, which is desired before data collection. This cannot be performed on gold grids.<\/p>\n

      It is important to plan\u00a0properly when working with gold grids. First,\u00a0load a carbon grid and align the microscope to a\u00a0perfect state, then load the gold grid for data collection.
      \n\"Overcoming<\/p>\n

      How many grids can be screened on the Tundra Cryo-TEM in a day?\u00a0\u00a0<\/em><\/strong><\/h2>\n

      Ten grids can be screened on the Tundra Cryo-TEM per day.\u00a0It takes 10 minutes to\u00a0load\u00a0a sample,\u00a020 minutes to\u00a0screen\u00a0one grid,\u00a0and\u00a0another 10 minutes to\u00a0unload.<\/p>\n

      It takes\u00a0in total\u00a040 minutes to screen one grid.<\/p>\n

      How often do I need to\u00a0bakeout\u00a0the sample loader and how long does it take?\u00a0<\/em><\/strong><\/h2>\n

      When working under cryo conditions, the cryo loading station needs to be baked out at least daily. There are two bakeout modes:<\/p>\n

        \n
      1. In-situ bakeout, which is\u00a0fully automated, and takes eight hours to complete. This can be used when the microscope is running\u00a0overnight.<\/li>\n
      2. Ex-situ mode, in which\u00a0user\u00a0need\u00a0to\u00a0empty the\u00a0liquid nitrogen pot manually. This takes roughly one hour and can be used during the day for a quick bakeout.<\/li>\n<\/ol>\n

        Are you doing demos, and can I send you samples?\u00a0\u00a0\u00a0<\/em><\/strong><\/h2>\n

        Yes, we are doing demos and will accommodate standard samples for analysis!<\/p>\n

        Contact us to learn more and schedule a demo >>\u00a0<\/a><\/p>\n

        View a special, 30-minute on-demand webinar for a closer look at our Tundra Cryo-TEM >><\/em><\/a><\/strong><\/p>\n

        \/\/<\/p>\n

        Lingbo Yu is a product marketing manager at Thermo Fisher Scientific.\u00a0<\/em><\/p>\n

        This post was written with the support of Dimple Karia, a product development engineer at Thermo Fisher Scientific.\u00a0<\/em><\/p>\n","protected":false},"excerpt":{"rendered":"

        At the dawn of 2021, we introduced a new, more accessible solution for cryo-electron microscopy: the Thermo Scientific Tundra Cryo-TEM. This smaller yet mighty cryo-EM instrument was designed to bring single particle analysis to every biochemistry laboratory. But what does that mean? You asked. We answered. Tundra Cryo-TEM: Commonly Asked Questions and Answers What type<\/p>\n","protected":false},"author":1519,"featured_media":635,"comment_status":"open","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"_acf_changed":false,"_kad_blocks_custom_css":"","_kad_blocks_head_custom_js":"","_kad_blocks_body_custom_js":"","_kad_blocks_footer_custom_js":"","_monsterinsights_skip_tracking":false,"_genesis_hide_title":false,"_genesis_hide_breadcrumbs":false,"_genesis_hide_singular_image":false,"_genesis_hide_footer_widgets":false,"_genesis_custom_body_class":"","_genesis_custom_post_class":"","_genesis_layout":"","_jetpack_newsletter_access":"","_jetpack_dont_email_post_to_subs":false,"_jetpack_newsletter_tier_id":0,"_jetpack_memberships_contains_paywalled_content":false,"_jetpack_feature_clip_id":0,"_jetpack_memberships_contains_paid_content":false,"footnotes":"","jetpack_post_was_ever_published":false},"categories":[24,33],"tags":[15,16,76,59],"division":[],"class_list":{"0":"post-610","1":"post","2":"type-post","3":"status-publish","4":"format-standard","5":"has-post-thumbnail","7":"category-cryo-electron-microscopy-cryo-em","8":"category-general","9":"tag-cryo-electron-microscopy","10":"tag-cryo-em","11":"tag-single-particle-analysis","12":"tag-transmission-electron-microscope","13":"entry"},"_selected_authors":"","_selected_reviewers":"","acf":[],"yoast_head":"\nCryo 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