Evaluation of hydrogen ion concentrations in prostates from rats and dogs using fluorescent confocal microscopy.
Evaluation of hydrogen ion concentrations in prostates from rats and dogs using fluorescent confocal microscopy.
复制标题
使用荧光共聚焦显微镜评估大鼠和狗前列腺中的氢离子浓度。
DOI：
10.1016/j.jphotobiol.2005.04.008
复制
发表时间：
2005
期刊：
影响因子：
--
通讯作者：
Levine,BarryS
中科院分区：
文献类型：
--
作者：
Lyubimov,AlexanderV;Carr,SerayaN;Brown,AlanP;Art,JonathanJ;Crowell,JamesA;Levine,BarryS
在反映人前列腺的动物模型中前列腺中pH的细胞内空间分布的知识可能对药物开发pH依赖性药物递送和活性具有影响。新鲜解剖的前列腺组织（体外）或整个前列腺（体内）加载荧光染料，并使用共聚焦显微镜观察。最初在用RPMI-1640培养基灌注的组织中拍摄图像。用含有尼日利亚菌素的已知pH的高钾缓冲液进行原位校准。乙酰氧基甲酯羧基-SNARF-1在大鼠和犬前列腺的上皮细胞（但非基质）中可见。通过Lyso Sensor绿色DND-189的荧光测定，前列腺上皮细胞中溶酶体的pH为1.52。通过二次加载和BCECF荧光染料的可视化开发了原位确认组织活力的方法。除了直接测量大鼠和犬组织中的pH值（pH ≥ 7.0）外，还开发了一种测量前列腺组织（而不是细胞培养物）中pH值的方法。
The knowledge of intracellular spatial distribution of pH in prostates in animal models reflective of human prostate may have implications for drug development upon pH dependent drug delivery and activity. Freshly dissected prostate tissues (in vitro) or the entire prostate gland (in vivo) were loaded with fluorescent dyes and viewed using confocal microscopy. Images were initially taken in tissues perfused with RPMI-1640 medium. Calibration in situ was performed with high potassium buffers of known pH containing nigericin. Acetoxymethyl ester carboxy-SNARF-1 was visible in epithelial cells (but not stroma) in rat and dog prostates. The pH of lysosomes in prostate epithelial cells was ⩽5.2 as determined by fluorescence of Lyso Sensor Green DND-189. A method of in situ confirmation of tissue viability was developed by a secondary loading and visualization of the BCECF fluorescent dye. Besides the direct measurement of the pH in rat and dog tissues (pH≈7.0), a method of pH measurement in prostate tissue (rather than in cell culture) was developed.