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Dataset . 2022
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ZENODO
Dataset . 2022
License: CC BY
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ZENODO
Dataset . 2022
License: CC BY
Data sources: Datacite
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Volumetric imaging of Arabidopsis thaliana root cells

Authors: Torres-Martínez, Héctor H.;

Volumetric imaging of Arabidopsis thaliana root cells

Abstract

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/m2s2 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.

{"references": ["Federici, F., Dupuy, L., Laplaze, L., Heisler, M., and Haseloff, J. Integrated genetic and computation methods for in planta cytometry, Nat. Methods 9, 483\u2013485 (2012).", "Geldner, N., D\u00e9nervaud-Tendon, V., Hyman, D. L., Mayer, U., Stierhof, Y.-D. and Chory, J. Rapid, combinatorial analysis of membrane compartments in intact plants with a multicolor marker set, The Plant J. 59, 169\u2013178 (2009).", "Schindelin, J. et al. Fiji: an open-source platform for biological-image analysis. Nat. Methods 9, 676\u2013682, (2012).", "Pitrone, P., Schindelin, J., Stuyvenberg, L. et al. OpenSPIM: an open-access light-sheet microscopy platform, Nat Methods 10, 598\u2013599 (2013).", "\"Download micro-manager latest release:\" https://micro-manager.org/Download_Micro-Manager_Latest_Release."]}

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popularity
This indicator reflects the "current" impact/attention (the "hype") of an article in the research community at large, based on the underlying citation network.
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influence
This indicator reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
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impulse
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