Direct visualization of Non-crystalline Cell wall Components by Ice-embedding Method.

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It became possible to observe aqueous suspension of cellulose directly with an electron microscopy. As the preliminary results of enzymatic hydrolysis of cellulose suggest that the efficiency of degradation can be influenced by the synergism of enzymes as well as the structure of cellulose, we planned to investigate the suspension with varying periods of reaction time. However, the problems such as the gap between the cooling fin of the anticontaminator did not fit the gap of the polepiece of the existing microscope, the astigmatism induced by the power control unit prevented the high resolution observation, took us considerable time to repair the entire system. To date, most of the problems, except for the image quality, has been overcome, and we could install the best condition (40 degree tilt) for the electron diffraction analyzes of cellulose. Therefore we slightly modified our initial planning in the way to determine the molecular directionality of cellulose molecules during biosynthesis, for instance, to determine whether cellulose is synthesized from the reducing end or non-reducing end sides. So far, remarkable progress has been made on this subject, and we could show the first experimental evidences that the reducing-end of cellulose molecule points outward the organism, in another word, the addition of monomer occurs at the non-reducing end of the preexisting cellulose molecule.