Development of confocal laser scanning micro-imaging analysis method of chloroplast photosynthetic functions in intact leaf

建立了完整叶片叶绿体光合作用功能的激光共聚焦扫描显微成像分析方法。对于用于CLSM系统的EM-CCD相机的A/D转换电平，在500到14,500的范围内，每个增益值的荧光强度与A/D转换电平之间显示出正的线性关系。激光功率与被测PPF之间的关系为线性方程。利用该系统对波士顿蕨类植物叶绿体的3D-F、3D-FM‘和3D-Φ图像进行了测量。还测量了茄子叶绿体向农化处理的暗/光转换(考茨基效应)。由于3DCLSM系统可以提供三维细胞排列中每个叶绿体的荧光强度、Φ、PSII和GT；以及NPQ值，因此该方法有可能在体内研究叶绿体功能的差异。

Confocal laser scanning micro-imaging(CLSM) analysis method of chloroplast photosynthetic functions in intact leaf was developed. For A/D conversion levels of EM-CCD camera used for the CLSM system the values revealed a positive linear relationship between fluorescence intensity and the A/D conversion level at each gain value in the range 500 to 14, 500. The relationship between laser power and measured PPF was expressed as a linear equation. Using this system, 3D-F, 3D-Fm' and 3D-Φ_<PSII> images of Boston fern chloroplasts were measured. The dark/light transitions(Kautsky effect) of eggplant chloroplasts to agrochemical treatment were also measured. As the 3D CLSM system can provide the fluorescence intensity,Φ_<PSII> and NPQ values of each chloroplast in a 3D cellular arrangement, this method has potential for investigating differences in the functions of chloroplasts in vivo.