摘要
探讨了CalciumOrange作为Harderian腺细胞胞浆Ca2 + 荧光指示探针的可能性 .发现选用 5 5 9nm作为激发光波长时 ,CalciumOrange有较强的吸收 ,而Harderian腺细胞 (含大量卟啉 )在此波长没有吸收 .利用CalciumOrange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CalciumOrange加载的Harderian腺细胞 ,Ionomycin引起荧光强度的增加 ,而用TritonX 10 0破裂细胞后加入钙离子螯合剂EGTA ,可使CalciumOrange荧光降到低于加入Ionomycin前的强度 .说明用 5 5 9nm为激发光波长 ,CalciumOrange可以用于Harderian腺细胞胞浆Ca2 + 的测定 .
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.
出处
《北京师范大学学报（自然科学版）》
CAS
CSCD
北大核心
2001年第5期675-679,共5页
Journal of Beijing Normal University(Natural Science)
基金
国家杰出青年科学基金资助项目 ( 3 982 5 112 )
国家自然科学基金资助项目 ( 3 9870 3 67
3 0 0 70 2 86)
