https://3demmethods.i2pc.es/index.php?action=history&feed=atom&title=2026Fromm_LowDose 2026-08-11T12:17:01Z Revision history for this page on the wiki MediaWiki 1.44.2 https://3demmethods.i2pc.es/index.php?title=2026Fromm_LowDose&diff=5242&oldid=prev 2026-08-11T05:28:04Z

<p>Created page with &quot;== Citation == Fromm, S.A. and Mattei, S. 2026. LowDoseWizard–rapid and standardised setup of low-dose cryo-TEM imaging in SerialEM. Acta Crystallographica Sec. D. 82, (2026), 990–997. == Abstract == Structure elucidation of biological macromolecules by single-particle cryogenic electron microscopy (SPA cryo-EM) or cryogenic electron tomography (cryo- ET) relies on low-dose imaging on cryogenic transmission electron microscopes (cryo-TEMs). Routine microscope setu...&quot;</p> <p><b>New page</b></p><div>== Citation ==<br /> <br /> Fromm, S.A. and Mattei, S. 2026. LowDoseWizard–rapid and standardised setup of low-dose cryo-TEM imaging in SerialEM. Acta Crystallographica Sec. D. 82, (2026), 990–997.<br /> <br /> == Abstract ==<br /> <br /> Structure elucidation of biological macromolecules by single-particle cryogenic<br /> electron microscopy (SPA cryo-EM) or cryogenic electron tomography (cryo-<br /> ET) relies on low-dose imaging on cryogenic transmission electron microscopes<br /> (cryo-TEMs). Routine microscope setup remains technically demanding and<br /> can be time-consuming, particularly for inexperienced or infrequent users. We<br /> present LowDoseWizard, a guided workflow implemented in SerialEM that<br /> enables rapid and standardized setup of cryo-TEM imaging conditions. From<br /> minimal user input, the workflow configures microscope optics, camera parameters<br /> and image-shift settings for all low-dose states, and guides the user<br /> through key daily alignment procedures, including beam-shift offset calibration,<br /> objective-lens astigmatism correction and coma-free alignment. The workflow<br /> is organized into modular routines that can be executed sequentially or independently,<br /> while microscope-specific acquisition parameters are defined in<br /> editable configuration files, allowing flexible adaptation to different instruments<br /> without modification of the core scripts. Across user sessions on three microscopes<br /> at EMBL Heidelberg, the complete setup required on average less than<br /> 15 min. To assess whether predefined imaging conditions generated by the<br /> workflow are compatible with high-resolution data collection, we acquired<br /> apoferritin data on a 200 kV Glacios and a 300 kV Titan Krios. These datasets<br /> yielded reconstructions at 1.62 and 1.09 A ˚ resolution, respectively, demonstrating<br /> that rapid, guided setup can support near-atomic and atomic-resolution<br /> SPA cryo-EM. LowDoseWizard lowers the barrier to robust cryo-TEM setup,<br /> reduces the time spent on routine parameter selection and alignment, and helps<br /> users focus on sample-specific aspects of data acquisition such as target selection.<br /> The workflow should be particularly valuable in shared instrumentation<br /> environments, where accessibility, reproducibility and efficient microscope use<br /> are critical.<br /> <br /> == Keywords ==<br /> <br /> == Links ==<br /> <br /> https://journals.iucr.org/d/issues/2026/08/00/sor5006/index.html<br /> <br /> == Related software ==<br /> <br /> == Related methods ==<br /> <br /> == Comments ==</div>

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