Microscope Imaging PAM Fluorometer

微型和大型藻类是水生生态系统功能的核心组成部分，提供初级生产和关键栖息地。藻类丰度或多样性的变化，特别是由于气候压力的增加，可能会对海洋食物网产生连锁影响。不幸的是，我们对藻类对环境胁迫的生理反应的了解通常仅限于宏观的海藻或浮游植物细胞在大量溶液中的研究，而我们对单个微观光养生物的生理知之甚少，包括光合繁殖体和对海藻繁殖周期至关重要的微观生命阶段。本申请申请的资金用于购买显微镜成像- pam（脉冲振幅调制）荧光仪，该荧光仪将独特地允许研究人员调查微观光合生物的压力生理，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）到微生物（例如，扁形虫），这些微生物吃了藻类并保留了可能或可能不仍然有效的叶绿体（kleptoplasty）。PAM荧光仪测量光养生物生理应激的光采集和叶绿素荧光；传统的PAM荧光仪使用光纤探针来传递光并检测溶液中宏观海藻或浮游植物细胞的荧光。这些传统的方法不足以检测单个细胞或单个微生物的荧光信号。所提出的显微镜成像- pam结合了传统的荧光测定法与显微镜和高灵敏度CCD相机和图像分析，以捕获显微光养体中叶绿素荧光的实时图像。这种创新的仪器同时记录光合参数和图像，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。这种新型设备将为研究海藻对环境胁迫的生命周期至关重要，为海藻养殖倡议和气候变化政策提供信息，以保护栖息地形成的大型藻类，也将为宿主微生物中kleptoplasids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具有广泛的应用和通用性，将大大拓宽微观光养生物生理实验的范围，并将加强HQP研究应激光生理的训练。

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.