High Throughput Fluorescence Imaging for Plant Sciences

植物科学高通量荧光成像

• 批准号: BB/R014086/1
• 负责人: Mark Fricker
• 金额: $ 37.45万
• 依托单位: University of Oxford
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2018
• 资助国家: 英国
• 起止时间: 2018 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FR014086%2F1
• 关键词: Throughput; Fluorescence; Imaging; Plant; Sciences

Research cell and developmental biology relies on imaging structures and molecules within living cells. In this way biological process can be followed in 3D over time scales ranging from tens of milliseconds to tens of hours. The most widely applied method for collecting such images is confocal fluorescence microscopy which provides clear images from within living tissues without the need to kill and physically section the specimen. The effectiveness of the method depends, first, on the ability to tag specific molecules with a fluorescent label and, second, on the ability of the microscope to form an image with high resolution and contrast whilst excluding out-of-focus information. Recent advances in structured illumination techniques, such as the Zeiss apotome system, have allowed optical sectioning at lower-magnifications which greatly facilitates initial fluorescence screening prior to confocal imaging, or measurements of physiological and developmental processes operating at much larger spatial scales (several cm squared) needed to image entire intact plant tissues and organs. Thus the ideal solution to be able to track developmental and physiological responses needs to combine low-magnification fluorescence for initial screening with high-resolution follow-up for detailed cellular and sub-cellular resolution. The addition of a robotic system tailored to the demands of plant specimens maximises the efficiency of data collection needed to achieve high-throughput for screening projects, or to allow long-term unattended operation for developmental studies. Such automated high-throughput fluorescence screening is routine in animal studies, where cells can be easily grown in multi-well culture plates, but a pipeline specifically tailored to handle the range of spatial scales and developmental time periods appropriate for plant systems would be unique in the UK.

研究细胞和发育生物学依赖于活细胞内的成像结构和分子。通过这种方式，可以在从几十毫秒到几十小时的时间范围内以3D方式跟踪生物过程。最广泛应用的收集这类图像的方法是共聚焦荧光显微镜，它从活组织内提供清晰的图像，而不需要杀死和物理切片标本。该方法的有效性首先取决于用荧光标记标记特定分子的能力，其次取决于显微镜在排除焦外信息的同时形成高分辨率和对比度图像的能力。结构照明技术的最新进展，如Zeiss apotome系统，允许在较低放大倍数下进行光学切片，这极大地促进了在共焦成像之前的初始荧光筛选，或在更大的空间尺度(几厘米的平方)上操作的生理和发育过程的测量，这是成像完整的植物组织和器官所必需的。因此，能够跟踪发育和生理反应的理想解决方案需要将用于初始筛查的低放大荧光与用于详细细胞和亚细胞分辨率的高分辨率后续检查相结合。增加一个为植物标本需求量身定做的机器人系统，最大限度地提高了数据收集的效率，以实现筛选项目的高通量，或允许长期无人值守的操作，以进行发育研究。这种自动化的高通量荧光筛选在动物研究中是例行公事，在动物研究中，细胞可以很容易地在多孔培养板中生长，但专门为处理适合植物系统的空间规模和发育时间范围的管道在英国将是独一无二的。

项目成果

A Novel Signaling Pathway Required for Arabidopsis Endodermal Root Organization Shapes the Rhizosphere Microbiome.

• DOI: 10.1093/pcp/pcaa170
• 发表时间: 2021-05-11
• 期刊: Plant & cell physiology
• 影响因子: 4.9
• 作者: Durr J;Reyt G;Spaepen S;Hilton S;Meehan C;Qi W;Kamiya T;Flis P;Dickinson HG;Feher A;Shivshankar U;Pavagadhi S;Swarup S;Salt D;Bending GD;Gutierrez-Marcos J
• 通讯作者: Gutierrez-Marcos J

A fundamental developmental transition in Physcomitrium patens is regulated by evolutionarily conserved mechanisms

• DOI: 10.1111/ede.12376
• 发表时间: 2021-04
• 期刊: Evolution & Development
• 影响因子: 2.9
• 作者: R. Jaeger;Laura A. Moody

Edge-based growth control in Arabidopsis involves two cell wall-associated Receptor-Like Proteins

• DOI: 10.1101/2022.06.10.495700
• 发表时间: 2022-06
• 期刊: bioRxiv
• 作者: Liam Elliott;M. Kalde;Ann-Kathrin Schuerholz;Sebastian Wolf;I. Moore;Charlotte Kirchhelle