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We have examined the method for the label-free optical microscopic image-based selection of cardiomyocytes from the mixture of collagenase-digested embryonic mouse heart cells using the on-chip imaging cell sorter system, in which 1/2000 s real-time high-speed camera-based phase-contrast cell image recognition was performed. To separate the cardiomyocytes from other cells including fibroblasts, we distinguished the roughness of the cell surface quantitatively as the index of sorting. The selected cells with smooth surface (roughness index not more than 1.1) for cardiomyocytes, and confirmed that the more than 80% of the collected and recultivated cells were self-beating and the 99.2% of them were identified as cardiomyocytes by immunostaining. The results indicates the potential of the label-free cell purification using imaging cell sorting system, especially for cardiomyocytes by the index of their cell surface roughness, which might be applicable for purification of cardiomyocytes for regenerative medicine. In conclusion, we have succeeded in separating cardiomyocytes with label-free on-chip imaging cell sorting procedures with using of the index of their surface roughness, and the results indicated that the more than 80% of the smooth cells were beating cardiomyocyte cells, and also more than 99% of healthy re-cultivated cells were cardiac myosin positive cells. The results suggested that the non-labeling simple phase-contrast image-based cell sorting has a potential to separate cardiomyocytes precisely from the mixture of heart cells by the index of cell surface roughness with less than 1 % error rate. [Nomura F, Kaneko T, Hattori A, Yasuda K (2011) Label-Free Shape-Based Selection of Cardiomyocytes with on-Chip Imaging Cell Sorting System]
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Last date updated on March, 2024

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