摘要
目的用钙的荧光指示剂ｆｌｕｏ－３和激光扫描共聚焦显微镜（ｌａｓｅｒｓｃａｎｎｉｎｇｃｏｎｆｏｃａｌｍｉｃｒｏｓｃｏｐｙ，ＬＳＣＭ）观察大白鼠的晶体上皮细胞（ｌｅｎｓｅｐｉｔｈｅｌｉａｌｃｅｌ，ＬＥＣ）中的游离钙。方法ｆｌｕｏ－３着染细胞后，应用ＬＳＣＭ观察细胞内ｆｌｕｏ－３与钙螯合后的荧光分布，最后用钙离子载体Ａ２３１８７和重金属离子Ｍｎ＋＋作校正，将ｆｌｕｏ－３与钙结合显示的荧光强度转换为［Ｃａ＋＋］ｉ值。结果ＬＥＣ中的游离钙主要分布在细胞核中。大白鼠晶体上皮细胞中的［Ｃａ＋＋］ｉ为２５９．７９±４９．２４ｎｍｏｌ／Ｌ。结论ｆｌｕｏ－３与ＬＥＣ孵育后能着染细胞。用ＬＳＣＭ能直观地观察细胞内钙的分布，并能通过校正得到细胞内游离钙的绝对值。这种成熟的方法。
Objective Using a new fluorescent Ca ++ indicator (fluo 3) and laser scanning confocal microscopy (LSCM) to monitor the intracellular Ca ++ of rat lens epithelial cell (LEC). Methods The LEC was loaded with fluo 3, then the distribution of free Ca ++ in the LEC was observed by using LSCM monitoring the distribution of fluorescence that is given due to the chelation between fluo 3 and Ca ++ . At last, ionophore (A 23187 ) and heavy metal ions (Mn ++ ) were applied to calibrate fluo 3 intensities to obtain [Ca ++ ] i value. Results The free calcium in LEC is mainly located in the nucleus. Free calcium concentration of rat LEC is 259.79±49.24 nmol/L. Conclusion Fluo 3 can be loaded into LEC by incubation with acetoxymethyl (AM) ester of the dye, and LSCM can be used to observe the cellular free calcium directly by surveying the fluorescence. This method will pave a way technically for the further study of intracellular free Ca ++ of the LEC.
出处
《中华眼科杂志》
CAS
CSCD
北大核心
1997年第4期297-300,共4页
Chinese Journal of Ophthalmology
关键词
晶体上皮细胞
钙
显微镜检
激光扫描
Lens Calcium Laser Microscopy examination, fluorescence
