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We have developed two-photon microscope which is optimized for in-vivo deep tissue imaging. We have developed two-photon microscope which is optimized for in-vivo deep tissue imaging. This system was characterized by signal decay and PSF measurement. Axial resolution of this system increases with increasing depth. And light scattering by tissue limits the imaging depth of two-photon microscope. Using commercial microscope body cannot detect all of emission light through the objective lens. So, we are upgrading two-photon microscope for detecting all of fluorescence signals which are ballistic and scattered. This new system provides deeper and better for in-vivo deep tissue imaging.
We present a combined high-speed two-photon microscopy (TPM) and Doppler optical coherence tomography (DOCT) which can provide complement information of in vivo tissues: molecular and cellular information with TPM and supracellular, tissue structure and vasculature with DOCT. This combined system can do simultaneous imaging in principle asusing separate light sources. Currently the combined system is under development, and preliminary data of individual imaging modalities are presented: fingertip and intralipid flowimaging in micro tube with DOCT, ex-vivo tissue imaging with TPM.
Information on the spatial distribution and organization of chloroplasts is very important to understand photosynthesis and other related mechanisms such as transpiration. During high irradiance, chloroplasts are mostly distributed on the upper side of the leave maximizing their light harvesting function but also protecting lower living tissues from photodamage. In order to monitor such movement effectively, we developed a two-photon microscopy (TPM) which allows to visualize in-vivo 3D imaging of chloroplasts. We monitored the change in organization and distribution of chloroplasts of maize plants grown at high radiation. During the successive imaging of the same area, the plants were kept in the darkroom. In addition, the change in chloroplasts size and distribution was compared between a young leaf and a senescent leaf. Such basic studies are expected to provide new insights in understanding photosynthesis efficiency by analyzing the change in distribution and organization of chloroplasts.