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共焦镜同时检测细胞内Ca2 和pH的变化 |
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黄行许 鲍永耀 黄辉 张薇 霍霞 朴英杰
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530 nm和大于605 nm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。
关键词 显微镜检查,荧光/方法;巨噬细胞/生理学;钙/分析;酸碱平衡;小鼠
中图号 R329.3
Simultaneous Detection of Free Cytosolic Calcium and Intracellular pH by Laser Scanning Confocal
Microscope Huang Xingxu, BaoYongyao, Huang Hui, et al.Central Laboratory, First Military Medical
University, 510515
Abstract 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 Ca2+-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 Ca2+ 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.
Key words microscopy,fluorescence/methods;macrophages/physiol;calcium/anal;acid-base equilibrium;
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