Photobleaching of Fluorescent Probe in Microfluorometry and Detection of Active Oxygen Species

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It is important to understand the cause of photobleaching and to know how to protect from the bleaching the fluorescent DNA probes stained cells from bleaching in microscopy. We examined the protective effect of sodium azide (NaN<SUB>3</SUB>) from photobleaching on the micrometer and the detection of active oxygen species produced in the probe solution by photoirradiation using an electron spin resonance (ESR) with a trapping agent. It was found that the bleaching of several probes was protected 11-99% by adding 1 mM NaN<SUB>3</SUB> as a scavenger for singlet oxygen. The NaN<SUB>3</SUB>-pro-tecting efficiency for photobleaching on the microscope corresponded to the typical ESR signal intensity with the spin trapping agent for singlet oxygen. The OH radical produced in the probe-photosensitization was also detected by the ESR measurement with the another trapping agent. From the results, it was found that there were two types (predominantly, singlet oxygen or radicals) of active oxygen species produced at the probe-photoreactions. It was considered that active oxygen species produced from these photoreactions oxidized the probes to decrease the fluorescence intensity of the DNA probes used in the microscopy, and that the NaN<SUB>3</SUB> protected the photobleaching considerably without selfquenching of fluorescence.

収録刊行物

  • Acta histochemica et cytochemica

    Acta histochemica et cytochemica 30(2), 189-191, 1997-04-01

    JAPAN SOCIETY OF HISTOCHEMISTRY AND CYTOCHEMISTRY

参考文献:  11件中 1-11件 を表示

被引用文献:  1件中 1-1件 を表示

各種コード

  • NII論文ID(NAID)
    10008606574
  • NII書誌ID(NCID)
    AA00508022
  • 本文言語コード
    ENG
  • 資料種別
    ART
  • ISSN
    00445991
  • データ提供元
    CJP書誌  CJP引用  J-STAGE 
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