温度对激光扫描共聚焦成像效果的影响

doi:10.7517/issn.1674-0475.180303

影像科学与光化学 ›› 2018, Vol. 36 ›› Issue (5): 443-452.DOI: 10.7517/issn.1674-0475.180303

温度对激光扫描共聚焦成像效果的影响

王娟娟, 魏学红

山西大学大型科学仪器中心, 山西太原 030006

收稿日期:2018-03-16 修回日期:2018-06-19 出版日期:2018-09-15 发布日期:2018-09-15
通讯作者: 王娟娟
基金资助:
2016年中央提升王娟娟人才事业启动经费（304545007）资助

The Effect of Temperature on Imaging of Confocal Laser Scanning Microscope

WANG Juanjuan, WEI Xuehong

Scientific Instrument Center, Shanxi University, Taiyuan 030006, Shanxi, P. R. China

Received:2018-03-16 Revised:2018-06-19 Online:2018-09-15 Published:2018-09-15

摘要/Abstract

摘要： 为了探讨活细胞工作站温度变化对激光扫描共聚焦显微镜（Confocal laser scanning microscope，CLSM）成像的影响，进行了升温和降温条件下铃兰草切片的动态图像采集实验，同时结合Zeiss LSM880的完美聚焦（Definite Focus）功能，采用局部荧光表达动态分析（Mean ROI），分别分析了升温和降温过程中图像的平均明场亮度及荧光强度。结果表明，使用Definite Focus功能前后，活细胞工作站升温和降温均引起了焦点漂移，影响了图像明场亮度和荧光强度，图像质量并未得到改善；活细胞工作站温度达到设定值后还会继续变化，需经过约20 min才能稳定，温度稳定后，焦点漂移现象可减弱，对图像明场亮度及荧光强度的影响也随之减小。因此，在激光扫描共聚焦显微镜成像过程中，一定要维持活细胞工作站温度及整个仪器工作环境温度的稳定，并同时采集明场图像，排除温度变化引起的焦点漂移及荧光强度变化，以便更好地提供准确的样品信号。

关键词: 激光扫描共聚焦显微镜, 温度, 焦点漂移, 明场亮度, 荧光强度

Abstract: In order to explore the effect of temperature on imaging of confocal laser scanning microscope (CLSM), dynamic image acquisition experiments of Convallaria Maiglöckchen slice with heating and cooling condition are designed and conducted combined with the definite focus of Zeiss LSM880. The average bright field brightness and fluorescence intensity of image in heating and cooling process were also analyzed by Mean ROI, respectively. The results indicated that before and after use of Definite Focus, the heating and cooling of the live cell workstation all caused the focus-drift, which greatly affected bright field brightness and fluorescence intensity, and the quality of image was not improved. The temperature of live cell workstation continued changed after the temperature reached the set point. It would take about 20 min to stabilize. Once the temperature was stable, the focus-shift was weakened, and the influence of temperature on the brightness and fluorescence intensity of image decreased. Therefore, it is necessary to maintain the stability of the temperature of live cell workstation and the working condition of instrument, acquire the bright field of image in the process of confocal laser scanning microscope imaging, so as to eliminate the focus-drift and fluorescence intensity change caused by the temperature change and provide the accurate sample signals.

Key words: confocal laser scanning microscopes (CLSM), temperature, focus-drift, bright field brightness, fluorescence intensity

王娟娟, 魏学红. 温度对激光扫描共聚焦成像效果的影响[J]. 影像科学与光化学, 2018, 36(5): 443-452.

WANG Juanjuan, WEI Xuehong. The Effect of Temperature on Imaging of Confocal Laser Scanning Microscope[J]. Imaging Science and Photochemistry, 2018, 36(5): 443-452.

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温度对激光扫描共聚焦成像效果的影响

The Effect of Temperature on Imaging of Confocal Laser Scanning Microscope

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