Microscope Imaging PAM Fluorometer

显微镜成像 PAM 荧光计

• 批准号: RTI-2021-00490
• 负责人: Martone, Patrick
• 金额: $ 6.59万
• 依托单位: University of British Columbia
• 依托单位国家: 加拿大
• 项目类别: Research Tools and Instruments
• 财政年份: 2020
• 资助国家: 加拿大
• 起止时间: 2020-01-01 至 2021-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=718840
• 关键词: Microscope; Imaging; PAM; Fluorometer

Micro- and macroalgae are central components in the functioning of aquatic ecosystems providing primary production and critical habitat. Shifts in algal abundance or diversity, particularly due to increasing climate stress, could have cascading impacts through marine food webs. Unfortunately, our understanding of the physiological responses of algae to environmental stress is generally limited to studies of macroscopic seaweeds or phytoplankton cells in bulk solution, and we know little about the physiology of individual microscopic phototrophs, including the photosynthetic propagules and microscopic life stages that are critical to seaweed reproductive cycles. Funding requested in this application is to acquire a Microscope Imaging-PAM (Pulse Amplitude Modulation) fluorometer, which will uniquely allow researchers to investigate the stress physiology of microscopic photosynthetic organisms, from single cells (e.g., unicellular algae, spores) and microscopic life stages of seaweeds (e.g., gametophytes, germlings) to microorganisms (e.g., flatworms) that have eaten algae and retained chloroplasts that may or may not still be functional (kleptoplasty'). PAM fluorometers measure light acquisition and chlorophyll fluorescence as proxies for physiological stress in phototrophs; conventional PAM fluorometers use fibre-optic probes to deliver light and detect fluorescence in macroscopic seaweeds or phytoplankton cells in solution. These conventional methods are insufficient to detect fluorescence signals from individual cells or single microorganisms. The proposed Microscope Imaging-PAM combines traditional fluorometry with microscopy and highly sensitive CCD cameras and image analysis to capture real-time images of chlorophyll fluorescence in microscopic phototrophs. This innovative instrument records photosynthetic parameters and images simultaneously, uniquely allowing users to differentiate the health of individual cells and chloroplasts within tissues and to link stress with changes in microorganism behaviour. This novel equipment will be essential to studies of seaweed life-cycles in response to environmental stress, informing seaweed aquaculture initiatives as well as climate change policy to preserve habitat-forming macroalgae, and will also create an exciting new research direction for the physiological study of kleptoplastids within host microorganisms. The Microscope Imaging-PAM has wide applications and versatility that will significantly broaden the scope of physiological experiments on microscopic phototrophs and will enhance the training of HQP studying stress photophysiology.

微型和大型藻类是提供初级生产和关键栖息地的水生生态系统功能的中心组成部分。藻类丰度或多样性的变化，特别是由于日益加剧的气候压力，可能会通过海洋食物网产生级联影响。不幸的是，我们对藻类对环境胁迫的生理反应的了解通常局限于对宏观海藻或浮游植物细胞的研究，而对单个微观光养生物的生理学知之甚少，包括对海藻繁殖周期至关重要的光合作用繁殖体和微观生命阶段。本申请申请所要求的资金将用于获得显微镜成像-PAM(脉冲幅度调制)荧光仪，这将独特地允许研究人员调查微观光合作用生物的应激生理学，从单细胞(例如单细胞藻类、孢子)和海藻的微观生命阶段(例如配子体、胚芽)到以海藻为食并保留可能或可能仍然具有功能的叶绿体的微生物(例如扁平线虫)。PAM荧光计测量光获取和叶绿素荧光，作为光养生物生理应激的替代指标；传统的PAM荧光计使用光纤探头传递光，并检测溶液中宏观海藻或浮游植物细胞的荧光。这些常规方法不足以检测来自单个细胞或单个微生物的荧光信号。提出的显微镜成像-PAM结合了传统的荧光与显微镜和高灵敏度的CCD相机和图像分析，以捕获实时图像的叶绿素荧光在微观光养生物。这种创新的仪器同时记录光合作用参数和图像，独特地允许用户区分组织内单个细胞和叶绿体的健康状况，并将压力与微生物行为的变化联系起来。这一新设备将对研究海藻对环境胁迫的生活周期、为海藻水产养殖计划和气候变化政策提供信息以保护栖息地形成的大型藻类至关重要，也将为宿主微生物内的klehop pltids的生理研究创造一个令人兴奋的新研究方向。显微镜成像-PAM具有广泛的适用性和通用性，将极大地拓宽微型光养生物的生理实验范围，并将加强HQP研究逆境光生理学的培训。