Measurement of intracellular oxygen levels using fluorescence lifetime imaging microscopy (FLIM)
Measurement of intracellular oxygen levels using fluorescence lifetime imaging microscopy (FLIM)
复制标题
使用荧光寿命成像显微镜 (FLIM) 测量细胞内氧水平
DOI：
10.1117/12.632827
复制
发表时间：
2005
期刊：
影响因子：
--
通讯作者：
M. Mycek
中科院分区：
文献类型：
--
作者：
Dhruv Sud;W. Zhong;D. Beer;M. Mycek
在生理温控条件下，通过监测氧敏感染料六水合二氯联吡啶(RTDP)的荧光寿命，在体内测量细胞内的氧水平。我们使用荧光寿命成像显微镜(FLIM)和一个独立的氧传感器来校准RTDP寿命随氧水平的相应变化。Flim方法重复性地量化了从(Barrett‘s)腺癌柱状细胞系(SEG-1)培养的活体人类细胞中的氧气水平。这些方法在研究3-D生物样品中的氧梯度和监测细胞代谢状态方面应该是有用的。
Intracellular oxygen levels were measured in vivo under physiological-temperature controlled conditions by monitoring the fluorescence lifetime of the oxygen sensitive dye ruthenium tris(2,2'-dipyridyl) dichloride hexahydrate (RTDP). We employed fluorescence lifetime imaging microscopy (FLIM) and an independent oxygen sensor to calibrate changes in RTDP lifetime with corresponding changes in oxygen level. The FLIM method reproducibly quantified oxygen levels in living human cells cultured from a (Barrett's) adenocarcinoma columnar cell line (SEG-1). Approaches such as those developed here should prove useful for studying oxygen gradients in 3-D biological specimens and for monitoring cellular metabolic status.