Volumetric imaging of Arabidopsis thaliana root cells
Arabidopsis thaliana seeds were surface sterilized, germinated and grown in Murashigue and Skoog medium at pH 5.7 and supplemented with vitamins (0.1 mg l-1 pyridoxine, 0.1 mg l-1 nicotinic acid), 0.8% agar, and 1% sucrose. Plants were grown at 21°C, 16/8-hour light/dark periods at 105 µmol/m 2 s 2...
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ftzenodo:oai:zenodo.org:6850745 2024-09-09T20:02:24+00:00 Volumetric imaging of Arabidopsis thaliana root cells Héctor H. Torres-Martínez Joseph G. Dubrovsky 2022-07-17 https://doi.org/10.5281/zenodo.6850745 eng eng Zenodo https://doi.org/10.5281/zenodo.6836636 https://doi.org/10.21203/rs.3.rs-984659/v1 https://doi.org/10.1101/2021.10.17.464398 https://doi.org/10.5281/zenodo.6850744 https://doi.org/10.5281/zenodo.6850745 oai:zenodo.org:6850745 info:eu-repo/semantics/openAccess Creative Commons Attribution 4.0 International https://creativecommons.org/licenses/by/4.0/legalcode info:eu-repo/semantics/other 2022 ftzenodo https://doi.org/10.5281/zenodo.685074510.5281/zenodo.683663610.21203/rs.3.rs-984659/v110.1101/2021.10.17.46439810.5281/zenodo.6850744 2024-07-26T21:23:06Z Arabidopsis thaliana seeds were surface sterilized, germinated and grown in Murashigue and Skoog medium at pH 5.7 and supplemented with vitamins (0.1 mg l-1 pyridoxine, 0.1 mg l-1 nicotinic acid), 0.8% agar, and 1% sucrose. Plants were grown at 21°C, 16/8-hour light/dark periods at 105 µmol/m 2 s 2 light intensity. Using a Zeiss Axiovert 200M microscope and a C-APO 63X, 1.2NA objective (Oberkochen, Germany), confocal volumetric imaging of the plant material was performed, with a pixel size of 404 nm and Z step size of 500 nm. Excitation of the sample was provided by a 488 nm laser, and a filter cube with 525/45 nm and 630/92 nm bandpass filters was used for yellow and red emission light collection, respectively. An inverted Olympus FV1000-IX81 confocal microscope equipped with a LUMFLN×60, 1.3NA S objective was used for root cell nuclear imaging. Sample excitation was achieved with a 543 nm laser and a BA560-660 filter was used for emission light collection. Pixel size was 41 nm, with a Z step size of 100 nm. A custom-built selective plane illumination microscopy (SPIM) system was used for imaging of primary root cells expressing p35s:H2B-R-RF. Sample excitation was achieved with a 561 nm laser using stroboscopic illumination. Emission was filtered via a multi-bandpass emission filter (Semrock, FF01-446/523/600/677-25 BrightLine). Volumetric imaging was done by mounting the sample on a four dimensional (XYZ, and Y rotation) motorized stage (Picard Industries). A sCMOS sensor (Hamamatsu, ORCA-Flash4.0 V2) was used for signal recording. The OpenSPIM plugin of µmanager (v.1.4 for windows) was used for control of acquisition parameters, sample translation and stroboscopic illumination. Collected images had a pixel size of 0.325µm, Z step size of 100 nm and a Y rotation step of 1.8°. Experimental procedures were approved by the Bioethics Committee of the Biotechnology Institute of the National Autonomous University of Mexico. Other/Unknown Material Orca Zenodo Olympus ENVELOPE(156.767,156.767,-80.217,-80.217) |
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English |
description |
Arabidopsis thaliana seeds were surface sterilized, germinated and grown in Murashigue and Skoog medium at pH 5.7 and supplemented with vitamins (0.1 mg l-1 pyridoxine, 0.1 mg l-1 nicotinic acid), 0.8% agar, and 1% sucrose. Plants were grown at 21°C, 16/8-hour light/dark periods at 105 µmol/m 2 s 2 light intensity. Using a Zeiss Axiovert 200M microscope and a C-APO 63X, 1.2NA objective (Oberkochen, Germany), confocal volumetric imaging of the plant material was performed, with a pixel size of 404 nm and Z step size of 500 nm. Excitation of the sample was provided by a 488 nm laser, and a filter cube with 525/45 nm and 630/92 nm bandpass filters was used for yellow and red emission light collection, respectively. An inverted Olympus FV1000-IX81 confocal microscope equipped with a LUMFLN×60, 1.3NA S objective was used for root cell nuclear imaging. Sample excitation was achieved with a 543 nm laser and a BA560-660 filter was used for emission light collection. Pixel size was 41 nm, with a Z step size of 100 nm. A custom-built selective plane illumination microscopy (SPIM) system was used for imaging of primary root cells expressing p35s:H2B-R-RF. Sample excitation was achieved with a 561 nm laser using stroboscopic illumination. Emission was filtered via a multi-bandpass emission filter (Semrock, FF01-446/523/600/677-25 BrightLine). Volumetric imaging was done by mounting the sample on a four dimensional (XYZ, and Y rotation) motorized stage (Picard Industries). A sCMOS sensor (Hamamatsu, ORCA-Flash4.0 V2) was used for signal recording. The OpenSPIM plugin of µmanager (v.1.4 for windows) was used for control of acquisition parameters, sample translation and stroboscopic illumination. Collected images had a pixel size of 0.325µm, Z step size of 100 nm and a Y rotation step of 1.8°. Experimental procedures were approved by the Bioethics Committee of the Biotechnology Institute of the National Autonomous University of Mexico. |
author2 |
Joseph G. Dubrovsky |
format |
Other/Unknown Material |
author |
Héctor H. Torres-Martínez |
spellingShingle |
Héctor H. Torres-Martínez Volumetric imaging of Arabidopsis thaliana root cells |
author_facet |
Héctor H. Torres-Martínez |
author_sort |
Héctor H. Torres-Martínez |
title |
Volumetric imaging of Arabidopsis thaliana root cells |
title_short |
Volumetric imaging of Arabidopsis thaliana root cells |
title_full |
Volumetric imaging of Arabidopsis thaliana root cells |
title_fullStr |
Volumetric imaging of Arabidopsis thaliana root cells |
title_full_unstemmed |
Volumetric imaging of Arabidopsis thaliana root cells |
title_sort |
volumetric imaging of arabidopsis thaliana root cells |
publisher |
Zenodo |
publishDate |
2022 |
url |
https://doi.org/10.5281/zenodo.6850745 |
long_lat |
ENVELOPE(156.767,156.767,-80.217,-80.217) |
geographic |
Olympus |
geographic_facet |
Olympus |
genre |
Orca |
genre_facet |
Orca |
op_relation |
https://doi.org/10.5281/zenodo.6836636 https://doi.org/10.21203/rs.3.rs-984659/v1 https://doi.org/10.1101/2021.10.17.464398 https://doi.org/10.5281/zenodo.6850744 https://doi.org/10.5281/zenodo.6850745 oai:zenodo.org:6850745 |
op_rights |
info:eu-repo/semantics/openAccess Creative Commons Attribution 4.0 International https://creativecommons.org/licenses/by/4.0/legalcode |
op_doi |
https://doi.org/10.5281/zenodo.685074510.5281/zenodo.683663610.21203/rs.3.rs-984659/v110.1101/2021.10.17.46439810.5281/zenodo.6850744 |
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