Study on adaptation of single photosynthetic cells against environmental stress by using three-dimensional super-resolution microscopy with time resolution ability
利用具有时间分辨率的三维超分辨显微镜研究单个光合细胞对环境胁迫的适应
基本信息
- 批准号:16072209
- 负责人:
- 金额:$ 16.19万
- 依托单位:
- 依托单位国家:日本
- 项目类别:Grant-in-Aid for Scientific Research on Priority Areas
- 财政年份:2004
- 资助国家:日本
- 起止时间:2004 至 2006
- 项目状态:已结题
- 来源:
- 关键词:
项目摘要
A near-infrared fs pulse was illuminated on a line in a specimen by a resonant scanning mirror oscillating at 7.9kHz, and total multiphoton-induced fluorescence from the linear region was focused on the slit of an imaging polychromator. An electron-multiplying CCD camera was used to resolve fluorescence of different colors at different horizontal pixels and fluorescence of different spatial positions in a specimen at different vertical pixels. The full widths at half maximum of the point-spread function of the system were estimated to be 0.39-0.40, 0.33 and 0.56-0.59μm for the X, Y and Z axes, respectively, at fluorescence wavelengths between 644 and 690nm. This microscope was applied to a study of the subcellular fluorescence spectra of thylakoid membranes in a cyanobacterium, Anabaena. It was found that the fluorescence intensity ratio between chlorophyll molecules mainly of photosystem II and phycobilin molecules of phycobilisome (chlorophyll/phycobilin), became lower as one probed deeper inside the cells. This was attributable not to position dependence of re-absorption or scattering effects, but to an intrinsic change in the local physiological state of the thylakoid membrane, with the help of a transmission spectral measurement of subcellular domains. Adaptation of the thylakoid membrane against light conditions, and its variation between chloroplasts and cyanobacteria, has been known for decades, but characterization based on fluorescence spectrum at a subcellular resolution has been scarce. Our newly developed spectroimaging system has also been applied to chloroplasts in tabacco, zea mays, and chlorella. Tissue dependence in the case of tobacco, daily cycle in the case of zea mays, and culture-condition dependence in the case of chlorella were found.
近红外飞秒脉冲通过振荡在7.9kHz的共振扫描镜照射在样品中的线上,并且来自线性区域的总多光子诱导荧光聚焦在成像多色仪的狭缝上。一个电子倍增CCD相机被用来解决不同颜色的荧光在不同的水平像素和荧光的不同空间位置的样品在不同的垂直像素。在644 ~ 690 nm的荧光波长范围内,系统的点扩散函数在X、Y和Z轴上的半峰宽分别为0.39-0.40、0.33和0.56-0.59μm。应用该显微镜对鱼腥藻类囊体膜的亚细胞荧光光谱进行了研究。发现主要是光系统II的叶绿素分子和藻胆体的藻胆素分子之间的荧光强度比(叶绿素/藻胆素)随着探测到细胞内更深而变低。这是由于不依赖于位置的再吸收或散射效应,但在类囊体膜的局部生理状态的内在变化,与亚细胞域的透射光谱测量的帮助。类囊体膜对光条件的适应,以及叶绿体和蓝藻之间的变化,已经知道了几十年,但基于亚细胞分辨率的荧光光谱的表征一直很少见。我们新开发的光谱成像系统也已应用于烟草、玉米和油菜的叶绿体。烟草的情况下,组织的依赖性,玉米的情况下,每日周期和培养条件的依赖性的情况下,小球藻被发现。
项目成果
期刊论文数量(107)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
Crystal structure of BchU, a methyltransferase involved in bacterio-chlorophyll c biosynthesis, and its complex with S-adenosylhomocystein : Implications for reaction mechanism
BchU(一种参与细菌叶绿素 C 生物合成的甲基转移酶)及其与 S-腺苷高半胱氨酸复合物的晶体结构:对反应机制的影响
- DOI:
- 发表时间:2006
- 期刊:
- 影响因子:0
- 作者:Kei Wada;Hitomi Yamaguchi;Jiro Harada;Keiko Niimi;Shigeaki Osumi;Hirozo Oh-oka;Hitoshi Tamiaki;Keiichi Fukuyama
- 通讯作者:Keiichi Fukuyama
ESR Signal of the Iron-sulfur Center FX and its Function in the Homodimeric Reaction Center of Heliobacterium Modesticaldum
普通幽门螺杆菌同二聚反应中心铁硫中心FX的ESR信号及其功能
- DOI:
- 发表时间:2006
- 期刊:
- 影响因子:0
- 作者:R. Miyamoto;et al.
- 通讯作者:et al.
Structural basis for the methylation mechanism in methyltransferase BchU involved in bacteriochlorophyll c biosynthesis
参与细菌叶绿素c生物合成的甲基转移酶BchU甲基化机制的结构基础
- DOI:
- 发表时间:2005
- 期刊:
- 影响因子:0
- 作者:K. Wada;et al.
- 通讯作者:et al.
フェムト秒テクノロジー 25章「フェムト秒レーザーによる光合成電子移動の多様性解明
飞秒技术第 25 章“利用飞秒激光器阐明光合电子转移的多样性”
- DOI:
- 发表时间:2006
- 期刊:
- 影响因子:0
- 作者:K.Sugawara;G.Hirabayashi;N.Kamiya;H.Kuramitz;S.Tanaka;熊崎茂一(1章のみ担当);熊崎茂一(分担執筆 1章分担当)
- 通讯作者:熊崎茂一(分担執筆 1章分担当)
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