دورية أكاديمية
Protocol for rapid clearing and staining of fixed Arabidopsis ovules for improved imaging by confocal laser scanning microscopy
| العنوان: | Protocol for rapid clearing and staining of fixed Arabidopsis ovules for improved imaging by confocal laser scanning microscopy |
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| المؤلفون: | Rachele Tofanelli, Athul Vijayan, Sebastian Scholz, Kay Schneitz |
| المصدر: | Plant Methods, Vol 15, Iss 1, Pp 1-13 (2019) |
| بيانات النشر: | BMC, 2019. |
| سنة النشر: | 2019 |
| المجموعة: | LCC:Plant culture LCC:Biology (General) |
| مصطلحات موضوعية: | 3D reconstruction, 3D organ models, Arabidopsis, ClearSee, Imaging, Ovule, Plant culture, SB1-1110, Biology (General), QH301-705.5 |
| الوصف: | Abstract Background A salient topic in developmental biology relates to the molecular and genetic mechanisms that underlie tissue morphogenesis. Modern quantitative approaches to this central question frequently involve digital cellular models of the organ or tissue under study. The ovules of the model species Arabidopsis thaliana have long been established as a model system for the study of organogenesis in plants. While ovule development in Arabidopsis can be followed by a variety of different imaging techniques, no experimental strategy presently exists that enables an easy and straightforward investigation of the morphology of internal tissues of the ovule with cellular resolution. Results We developed a protocol for rapid and robust confocal microscopy of fixed Arabidopsis ovules of all stages. The method combines clearing of fixed ovules in ClearSee solution with marking the cell outline using the cell wall stain SCRI Renaissance 2200 and the nuclei with the stain TO-PRO-3 iodide. We further improved the microscopy by employing a homogenous immersion system aimed at minimizing refractive index differences. The method allows complete inspection of the cellular architecture even deep within the ovule. Using the new protocol we were able to generate digital three-dimensional models of ovules of various stages. Conclusions The protocol enables the quick and reproducible imaging of fixed Arabidopsis ovules of all developmental stages. From the imaging data three-dimensional digital ovule models with cellular resolution can be rapidly generated using image analysis software, for example MorphographX. Such digital models will provide the foundation for a future quantitative analysis of ovule morphogenesis in a model species. |
| نوع الوثيقة: | article |
| وصف الملف: | electronic resource |
| اللغة: | English |
| تدمد: | 1746-4811 |
| Relation: | http://link.springer.com/article/10.1186/s13007-019-0505-x; https://doaj.org/toc/1746-4811 |
| DOI: | 10.1186/s13007-019-0505-x |
| URL الوصول: | https://doaj.org/article/709b3e89c10b4af9baeca6aa5a4ad997 |
| رقم الأكسشن: | edsdoj.709b3e89c10b4af9baeca6aa5a4ad997 |
| قاعدة البيانات: | Directory of Open Access Journals |
Find this article in full text from Springer Verlag. 
Full Text Finder | تدمد: | 17464811 |
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| DOI: | 10.1186/s13007-019-0505-x |