• Chinese Journal of Lasers
  • Vol. 45, Issue 4, 407001 (2018)
Xu Yiwen1、2, Zhang Yunhai1、*, Yang Haomin1, Ji Lin1, Chang Jian1, Liu Chuang1、2, and Tang Yuguo1
Author Affiliations
  • 1[in Chinese]
  • 2[in Chinese]
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    DOI: 10.3788/CJL201845.0407001 Cite this Article Set citation alerts
    Xu Yiwen, Zhang Yunhai, Yang Haomin, Ji Lin, Chang Jian, Liu Chuang, Tang Yuguo. Imaging Technology of Reduced Photobleaching Based on Controllable Light Exposure-Confocal Microscopy[J]. Chinese Journal of Lasers, 2018, 45(4): 407001 Copy Citation Text show less

    Abstract

    Photobleaching induced by excitation light limits the application of confocal microscopy for long-time observation of biological samples. A new imaging approach, controllable light exposure-confocal microscopy (CLE-CM), is proposed. CLE-CM has two thresholds decided through pre-experiments and detects the feedback of sample pixel value at set intervals. The feedback is compared with upper and lower thresholds. By spatially controlling the light-exposure time of objective pixel according to the results of comparison, CLE-CM exploits fluorescent information in a more efficient way and reduces photobleaching without decreasing image quality. Two scan series of 11 successive CLE-CM images and standard confocal images show that, CLE-CM reduces 52.62% of photobleaching in bovine pulmonary artery endothelial (BPAE) cells compared with standard confocal at the 11th image. The effect of bleaching attenuation depends on the fluorophore distribution. CLE-CM decreases photobleaching markedly through the reduction of excitation-light dose, and increases the number of good-quality images that can be captured continuously by confocal microscope.
    Xu Yiwen, Zhang Yunhai, Yang Haomin, Ji Lin, Chang Jian, Liu Chuang, Tang Yuguo. Imaging Technology of Reduced Photobleaching Based on Controllable Light Exposure-Confocal Microscopy[J]. Chinese Journal of Lasers, 2018, 45(4): 407001
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