Imaging Science and Photochemistry ›› 2018, Vol. 36 ›› Issue (5): 443-452.DOI: 10.7517/issn.1674-0475.180303

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The Effect of Temperature on Imaging of Confocal Laser Scanning Microscope

WANG Juanjuan, WEI Xuehong   

  1. 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

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