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

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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.