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cryoWriter: a blotting free cryo-EM preparation system with a climate jet and cover-slip injector
Electron microscopy (EM) introduced a fast and lasting change to structural and cellular biology. However, the sample preparation is still the bottleneck in the cryogenic electron microscopy (cryo-EM) workflow. Classical specimen preparation methods employ a harsh paper-blotting step, and the protei...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
The Royal Society of Chemistry
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9641993/ https://www.ncbi.nlm.nih.gov/pubmed/35924676 http://dx.doi.org/10.1039/d2fd00066k |
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author | Rima, Luca Zimmermann, Michael Fränkl, Andri Clairfeuille, Thomas Lauer, Matthias Engel, Andreas Engel, Hans-Andreas Braun, Thomas |
author_facet | Rima, Luca Zimmermann, Michael Fränkl, Andri Clairfeuille, Thomas Lauer, Matthias Engel, Andreas Engel, Hans-Andreas Braun, Thomas |
author_sort | Rima, Luca |
collection | PubMed |
description | Electron microscopy (EM) introduced a fast and lasting change to structural and cellular biology. However, the sample preparation is still the bottleneck in the cryogenic electron microscopy (cryo-EM) workflow. Classical specimen preparation methods employ a harsh paper-blotting step, and the protein particles are exposed to a damaging air–water interface. Therefore, improved preparation strategies are urgently needed. Here, we present an amended microfluidic sample preparation method, which entirely avoids paper blotting and allows the passivation of the air–water interface during the preparation process. First, a climate jet excludes oxygen from the sample environment and controls the preparation temperature by varying the relative humidity of the grid environment. Second, the integrated “coverslip injector” allows the modulation of the air–water interface of the thin sample layer with effector molecules. We will briefly discuss the climate jet’s effect on the stability and dynamics of the sample thin films. Furthermore, we will address the coverslip injector and demonstrate significant improvement in the sample quality. |
format | Online Article Text |
id | pubmed-9641993 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | The Royal Society of Chemistry |
record_format | MEDLINE/PubMed |
spelling | pubmed-96419932022-11-14 cryoWriter: a blotting free cryo-EM preparation system with a climate jet and cover-slip injector Rima, Luca Zimmermann, Michael Fränkl, Andri Clairfeuille, Thomas Lauer, Matthias Engel, Andreas Engel, Hans-Andreas Braun, Thomas Faraday Discuss Chemistry Electron microscopy (EM) introduced a fast and lasting change to structural and cellular biology. However, the sample preparation is still the bottleneck in the cryogenic electron microscopy (cryo-EM) workflow. Classical specimen preparation methods employ a harsh paper-blotting step, and the protein particles are exposed to a damaging air–water interface. Therefore, improved preparation strategies are urgently needed. Here, we present an amended microfluidic sample preparation method, which entirely avoids paper blotting and allows the passivation of the air–water interface during the preparation process. First, a climate jet excludes oxygen from the sample environment and controls the preparation temperature by varying the relative humidity of the grid environment. Second, the integrated “coverslip injector” allows the modulation of the air–water interface of the thin sample layer with effector molecules. We will briefly discuss the climate jet’s effect on the stability and dynamics of the sample thin films. Furthermore, we will address the coverslip injector and demonstrate significant improvement in the sample quality. The Royal Society of Chemistry 2022-08-04 /pmc/articles/PMC9641993/ /pubmed/35924676 http://dx.doi.org/10.1039/d2fd00066k Text en This journal is © The Royal Society of Chemistry https://creativecommons.org/licenses/by/3.0/ |
spellingShingle | Chemistry Rima, Luca Zimmermann, Michael Fränkl, Andri Clairfeuille, Thomas Lauer, Matthias Engel, Andreas Engel, Hans-Andreas Braun, Thomas cryoWriter: a blotting free cryo-EM preparation system with a climate jet and cover-slip injector |
title | cryoWriter: a blotting free cryo-EM preparation system with a climate jet and cover-slip injector |
title_full | cryoWriter: a blotting free cryo-EM preparation system with a climate jet and cover-slip injector |
title_fullStr | cryoWriter: a blotting free cryo-EM preparation system with a climate jet and cover-slip injector |
title_full_unstemmed | cryoWriter: a blotting free cryo-EM preparation system with a climate jet and cover-slip injector |
title_short | cryoWriter: a blotting free cryo-EM preparation system with a climate jet and cover-slip injector |
title_sort | cryowriter: a blotting free cryo-em preparation system with a climate jet and cover-slip injector |
topic | Chemistry |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9641993/ https://www.ncbi.nlm.nih.gov/pubmed/35924676 http://dx.doi.org/10.1039/d2fd00066k |
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