| 利用Calcium Orange测定Harderian腺细胞中的钙离子浓度 |
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| 引用本文： | 周衍东,刘玉芝,崔宗杰.利用Calcium Orange测定Harderian腺细胞中的钙离子浓度[J].北京师范大学学报(自然科学版),2001,37(5):675-679. |
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| 作者姓名： | 周衍东 刘玉芝 崔宗杰 |
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| 作者单位： | 1. 北京师范大学细胞生物学研究所
2. 河北农业大学动物科技学院 |
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| 基金项目： | 国家自然科学基金;39825112; |
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| 摘 要： | 探讨了Calcium Orange作为Harderian腺细胞胞浆Ca^2 荧光指示探针的可能性，发现选用559nm作为激发光波长时，Calcium Orange有较强制 吸收，而Harderian腺细胞（含大量卟啉）在此波长没有吸收，利用Calcium Orange作为指示剂可以避免紫外光所引发的原卟啉介导的光动力作用，在Calcium Orange加载的Harderian腺细胞，Ionomycin引起荧光强度的增加，而用TritonX-100破裂细胞后加入钙离子螯合剂EGTA，可使Calcium Orange荧光降到低于加入Ionomycin前的强度，说明用559nm为激发光波长，Calcium Orange可以用于Harderian腺细胞胞浆Ca^2 的测定。
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| 关 键 词： | Harderian腺细胞 原卟啉Ⅸ CalciumOrange fura－2 光动力作用 钙离子浓度 眼球 |
| 修稿时间： | 2001年6月7日 |
MEASUREMENT OF INTRACELLULAR CALCIUM CONCENTRATION IN THE ISOLATED RAT HARDERIAN GLAND CELLS USING CALCIUM ORANGE |
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| Zhou Yandong,Liu Yuzhi,Cui Zongjie.MEASUREMENT OF INTRACELLULAR CALCIUM CONCENTRATION IN THE ISOLATED RAT HARDERIAN GLAND CELLS USING CALCIUM ORANGE[J].Journal of Beijing Normal University(Natural Science),2001,37(5):675-679. |
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| Authors: | Zhou Yandong Liu Yuzhi Cui Zongjie |
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| Institution: | Zhou Yandong 1) Liu Yuzhi 2) Cui Zongjie 1) |
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| Abstract: | The use of Calcium Orange to measure intracellular calcium concentration in the isolated rat Harderian gland cells is investigated. After comparing the excitation spectra, it is found that Calcium Orange loaded in duck pancreatic acini has strong absorption at 559 nm, whereas Harderian gland cells have no absorption at this wavelength. Therefore, light at 559 nm will not trigger protoporphyrin-mediated photodynamic action. In Calcium Orange loaded Harderian gland cells, Ionomycin induces significant increase in fluorescence, and after cell lysis with Triton X-100, addition of EGTA diminishes the fluorescence to below the level before addition of Ionomycin. In conclusion, Calcium Orange can be used as a calcium indicator in the isolated Harderian gland cells with the excitation wavelength at 559 nm. |
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| Keywords: | CalciumOrange fura-2 |
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