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，可能通过海洋食物网产生连锁反应。不幸的是，我们对藻类对环境胁迫的生理反应的理解通常仅限于宏观海藻或浮游植物细胞在散装溶液中的研究，我们对个体微观光养生物的生理学知之甚少，包括光合繁殖体和微观生命阶段，这对海藻繁殖周期至关重要。该申请中要求的资金是获得显微镜成像-PAM（脉冲幅度调制）荧光计，这将使研究人员能够从单细胞（例如，单细胞藻类，孢子）和海藻的微观生命阶段（例如，配子体，幼苗）到微生物（例如，扁形虫），它们吃了藻类，并保留了可能仍有功能或可能没有功能的叶绿体（“盗质体”）。PAM荧光计测量光采集和叶绿素荧光作为光养生物生理应激的代理;传统的PAM荧光计使用光纤探针来提供光并检测溶液中宏观海藻或浮游植物细胞中的荧光。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。建议显微镜成像PAM结合传统的荧光显微镜和高灵敏度的CCD相机和图像分析，捕捉实时图像的叶绿素荧光显微光养生物。这款创新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。这种新型设备将是必不可少的海藻生命周期的研究，以应对环境压力，通知海藻水产养殖计划以及气候变化政策，以保护栖息地形成的大型藻类，也将创造一个令人兴奋的新的研究方向的kleptoplastids在宿主微生物的生理研究。显微成像PAM具有广泛的应用性和通用性，将大大拓宽微观光养生物生理实验的范围，并将加强HQP研究胁迫生理学的培训。

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.