Autofluorescence lifetime imaging of cultivated cells using a UV picosecond laser diode
Autofluorescence lifetime imaging of cultivated cells using a UV picosecond laser diode
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DOI：
10.1023/b:jofl.0000039351.09916.cc
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发表时间：
2004-09-01
影响因子：
2.7
通讯作者：
Sailer, R
中科院分区：
文献类型：
--
作者：
Schneckenburger, H;Wagner, M;Sailer, R
使用一种新型的皮秒激光二极管在近紫外范围（375 nm）的培养内皮细胞的自发荧光的寿命图像记录。与现有的皮秒光源相比，该波长允许分别具有0.4-0.5 ns和2.0-2.5 ns的荧光寿命的游离和蛋白质结合的辅酶NADH的有效激发。有效荧光寿命τ（eff）（取决于两个寿命）均匀分布在细胞核周区域，可能接近线粒体的一些缩短。在线粒体呼吸链抑制后观察到画眉草的轻微减少，而在糖酵解途径抑制后观察到画眉草的轻微增加，从而表明游离和蛋白质结合的NADH的比率的变化。虽然目前的应用仍然受到其低脉冲能量（小于或等于5 pJ）的限制，紫外皮秒激光二极管在高分辨率荧光显微镜和荧光寿命内窥镜中具有很大的潜力。
Lifetime images of autofluorescence of cultivated endothelial cells were recorded using a novel picosecond laser diode in the near ultraviolet range (375 nm). In contrast to existing picosecond light sources this wavelength permits efficient excitation of the free and protein bound coenzyme NADH with fluorescence lifetimes of 0.4-0.5 ns and 2.0-2.5 ns, respectively. The effective fluorescence lifetime tau(eff) (depending on both lifetimes) was homogenously distributed over the cells with some shortening in the perinuclear region, possibly close to mitochondria. A slight decrease of teff was observed after inhibition of the mitochondrial respiratory chain, whereas a slight increase was observed after inhibition of the glycolytic pathway, thus indicating variations of the ratio of free and protein bound NADH. Although present applications are still limited by their low pulse energy (less than or equal to5 pJ), uv picosecond laser diodes have a large potential in high resolution fluorescence microscopy and fluorescence lifetime endoscopy.