Grid batch-dependent tuning of glow discharge parameters
Sample preparation on cryo-EM grids can give various results, from very thin ice and homogeneous particle distribution (ideal case) to unwanted behavior such as particles around the “holes” or complexes that do not entirely correspond to the one in solution (real life). We recently run into such a c...
| Main Authors: | , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Frontiers Media S.A.
2022-08-01
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| Series: | Frontiers in Molecular Biosciences |
| Subjects: | |
| Online Access: | https://www.frontiersin.org/articles/10.3389/fmolb.2022.910218/full |
| _version_ | 1798042329591840768 |
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| author | Ramy Kazan Gabrielle Bourgeois Dominique Carisetti Ileana Florea Eric Larquet Jean-Luc Maurice Yves Mechulam François Ozanam Emmanuelle Schmitt Pierre-Damien Coureux |
| author_facet | Ramy Kazan Gabrielle Bourgeois Dominique Carisetti Ileana Florea Eric Larquet Jean-Luc Maurice Yves Mechulam François Ozanam Emmanuelle Schmitt Pierre-Damien Coureux |
| author_sort | Ramy Kazan |
| collection | DOAJ |
| description | Sample preparation on cryo-EM grids can give various results, from very thin ice and homogeneous particle distribution (ideal case) to unwanted behavior such as particles around the “holes” or complexes that do not entirely correspond to the one in solution (real life). We recently run into such a case and finally found out that variations in the 3D reconstructions were systematically correlated with the grid batches that were used. We report the use of several techniques to investigate the grids' characteristics, namely TEM, SEM, Auger spectroscopy and Infrared Interferometry. This allowed us to diagnose the origin of grid preparation problems and to adjust glow discharge parameters. The methods used for each approach are described and the results obtained on a common specific case are reported. |
| first_indexed | 2024-04-11T22:34:03Z |
| format | Article |
| id | doaj.art-980ec9147ab44014acdc832a74586804 |
| institution | Directory Open Access Journal |
| issn | 2296-889X |
| language | English |
| last_indexed | 2024-04-11T22:34:03Z |
| publishDate | 2022-08-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Molecular Biosciences |
| spelling | doaj.art-980ec9147ab44014acdc832a745868042022-12-22T03:59:16ZengFrontiers Media S.A.Frontiers in Molecular Biosciences2296-889X2022-08-01910.3389/fmolb.2022.910218910218Grid batch-dependent tuning of glow discharge parametersRamy Kazan0Gabrielle Bourgeois1Dominique Carisetti2Ileana Florea3Eric Larquet4Jean-Luc Maurice5Yves Mechulam6François Ozanam7Emmanuelle Schmitt8Pierre-Damien Coureux9Laboratoire de Biologie Structurale de la Cellule, BIOC, Ecole Polytechnique, CNRS-UMR7654, IP Paris, Palaiseau, FranceLaboratoire de Biologie Structurale de la Cellule, BIOC, Ecole Polytechnique, CNRS-UMR7654, IP Paris, Palaiseau, FranceThales Research and Technology, Palaiseau, FranceLaboratoire de Physique des Interfaces et Couches Minces (LPICM), CNRS-UMR 7647, Ecole Polytechnique, IP Paris, Palaiseau, FranceLaboratoire de Physique de la Matière Condensée, Ecole Polytechnique, CNRS, IP Paris, Palaiseau, FranceLaboratoire de Physique des Interfaces et Couches Minces (LPICM), CNRS-UMR 7647, Ecole Polytechnique, IP Paris, Palaiseau, FranceLaboratoire de Biologie Structurale de la Cellule, BIOC, Ecole Polytechnique, CNRS-UMR7654, IP Paris, Palaiseau, FranceLaboratoire de Physique de la Matière Condensée, Ecole Polytechnique, CNRS, IP Paris, Palaiseau, FranceLaboratoire de Biologie Structurale de la Cellule, BIOC, Ecole Polytechnique, CNRS-UMR7654, IP Paris, Palaiseau, FranceLaboratoire de Biologie Structurale de la Cellule, BIOC, Ecole Polytechnique, CNRS-UMR7654, IP Paris, Palaiseau, FranceSample preparation on cryo-EM grids can give various results, from very thin ice and homogeneous particle distribution (ideal case) to unwanted behavior such as particles around the “holes” or complexes that do not entirely correspond to the one in solution (real life). We recently run into such a case and finally found out that variations in the 3D reconstructions were systematically correlated with the grid batches that were used. We report the use of several techniques to investigate the grids' characteristics, namely TEM, SEM, Auger spectroscopy and Infrared Interferometry. This allowed us to diagnose the origin of grid preparation problems and to adjust glow discharge parameters. The methods used for each approach are described and the results obtained on a common specific case are reported.https://www.frontiersin.org/articles/10.3389/fmolb.2022.910218/fullcryo-EMsample preparationglow dischargequantifoilcontinuous carbon |
| spellingShingle | Ramy Kazan Gabrielle Bourgeois Dominique Carisetti Ileana Florea Eric Larquet Jean-Luc Maurice Yves Mechulam François Ozanam Emmanuelle Schmitt Pierre-Damien Coureux Grid batch-dependent tuning of glow discharge parameters Frontiers in Molecular Biosciences cryo-EM sample preparation glow discharge quantifoil continuous carbon |
| title | Grid batch-dependent tuning of glow discharge parameters |
| title_full | Grid batch-dependent tuning of glow discharge parameters |
| title_fullStr | Grid batch-dependent tuning of glow discharge parameters |
| title_full_unstemmed | Grid batch-dependent tuning of glow discharge parameters |
| title_short | Grid batch-dependent tuning of glow discharge parameters |
| title_sort | grid batch dependent tuning of glow discharge parameters |
| topic | cryo-EM sample preparation glow discharge quantifoil continuous carbon |
| url | https://www.frontiersin.org/articles/10.3389/fmolb.2022.910218/full |
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