共焦镜同时检测细胞内Ca^(2+)和pH的变化  

Simultaneous Detection of Free Cytosolic Calcium and Intracellular pH by Laser Scanning Confocal Microscope

作　　者：黄行许[1] 鲍永耀[1] 黄辉[1] 张薇[1] 霍霞[1] 朴英杰[1] 

机构地区：[1]第一军医大学中心实验室

出　　处：《中华物理医学杂志》1998年第1期35-37,共3页Chinese Journal of Physical Medicine and Rehabilitation

摘　　要：目的细胞内游离Ｃａ２＋与胞浆的ｐＨ在不同因素作用下可以改变。研究可以同时检测游离Ｃａ２＋与ｐＨ变化的方法激光扫描共聚焦显微镜观察。方法用胞内游离Ｃａ２＋和ｐＨ荧光探针ｆｌｕｏ３／ＡＭ和ＳＮＡＲＦ１／ＡＭ标记小鼠腹腔巨噬细胞后，在激光扫描共聚焦显微镜（ＡＣＡＳ５７０）下，同时用最适发射波长分别为５３０ｎｍ和大于６０５ｎｍ的检测器１和检测器２检测巨噬细胞内ｆｌｕｏ３／ＡＭ和ＳＮＡＲＦ１／ＡＭ的荧光强度变化。结果检测器１和检测器２分别只检测反映细胞内游离Ｃａ２＋和ｐＨ变化的ｆｌｕｏ３／ＡＭ和ＳＮＡＲＦ１／ＡＭ所发荧光的变化；ＳＮＡＲＦ１／ＡＭ和ｆｌｕｏ３／ＡＭ所发荧光相互不影响。结论激光扫描共聚焦显微镜可同时检测细胞内游离Ｃａ２＋和ｐＨ的变化。Objective Free cytosolic calcium and intracellular pH can have some changes due to certain factors.It is to explore a method for simultaneous detection on the changes,that is,observation with laser scanning confocal microscope.Methods Ca2+and pHsensitive fluorescence probes of fluo3/AM and SNARF1/AM were used to mark the peritoneal macrophages simultaneously. Then, the variations of the fluorescence were detected with detector 1 (optimum emission wavelength: 530nm) and detector 2 (optimum emission wavelength: >605nm) of the laser scanning confocal microscope (ACAS570). Results Detector 1 and 2 each detected the variations of the fluorescence of fluo3/Am or SNARF1/AM of the Ca2+ or pH. The fluorescence of fluo3/AM and SNARF1/AM did not influence each other. And the forms of the alteration in free cytosolic calcium and intracellular pH of the macrophages varied when they were stimulated by different stimulators. Conclusion Laser scanning confocal microscope could be used in simultaneous detection of free cytosolic calcium and intracellular pH.

关 键 词：显微镜检查 共焦镜 钙 PH值 

分 类 号：R329.26[医药卫生—人体解剖和组织胚胎学]