TED-AJ03-367 INFLUENCE OF CRYOPROTECTANT ON TIME-SERIES THREE-DIMENSIONAL MICROSTRUCTURE OF BIOLOGICAL TISSUES DURING FREEZING AND THAWING
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概要
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Freezing and thawing of biological tissues are fundamental phenomena in cryopreservation and cryosurgery. The freezing and thawing of the tissues generally proceed transiently and spatially in three-dimensions(3D). Therefore, 3D visualization in real time is required to understand the details of microstructure in the process. Three-dimensional behavior of ice crystals and cells during the freezing and thawing of biological tissues was investigated microscopically in real time by using a confocal laser scanning microscope (CLSM) and a fluorescent dye to clarify the influence of cryoprotectant on the time-series 3D microstructure. Experimental materials were fresh tender meat (2nd pectoral muscles) of chicken protected with 2.0M dimethyl sulfoxide (DMSO), a common cryoprotectant penetrating cells, in physiological saline. The tissues was stained with a fluoresent dye, acridine orange, to distinguish ice crystals and cells (muscle fibers) by their different colors using the CLSM/dye method. Then, the tissues were frozen and thawed in a uniform temperature on the cryostage (Figure A-1) under two different thermal protocols : a)slow-cooling (H=1.0℃/min) and rapid-warming (W∿100℃/min) and b)rapid-cooling (W∿100℃/min) and rapid-warming (W∿100℃/min). The CLSM noninvasively produced tomograms of the tissues to clarify the pattern of freezing, morphology of extra- and intracellular ice crystals in the tissues, and the interaction between ice crystals and cells. (Figure A-2 shows an example of visualization. IC(ex) and MF respectively denote extracellular ice crystals and muscle fibers.) The results were compared between the thermal protocols, and comapred with the microstructure without DMSO. After freezing and thawing, the tissues was also investigated by histological means. The histological changes of tissues were correlated with the results from time-series 3D visualization.[figure]
- 一般社団法人日本機械学会の論文
著者
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Ishiguro Hiroshi
Department Of Gastroenterology Nagoya University Graduate School Of Medicine
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Ishiguro Hiroshi
Department Of Adaptive Machine Systems Osaka University
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Ishiguro Hiroshi
Department Of Biological Functions And Engineering Graduate School Of Life Science And Systems Engin
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HORIMIZU Takashi
Graduate School of Engineering Mechanics, University of Tsukuba
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KATAORI Akinobu
Graduate School of Engineering Mechanics, University of Tsukuba
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Kataori Akinobu
Graduate School Of Engineering Mechanics University Of Tsukuba
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Horimizu Takashi
Graduate School Of Engineering Mechanics University Of Tsukuba
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Ishiguro Hiroshi
Department Of Adaptive Machine Systems Graduate School Of Engineering Osaka University
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