Fluorescence lifetime measurements in confocal microscopy of neurons labeled with multiple fluorophores
Fluorescence lifetime measurements in confocal microscopy of neurons labeled with multiple fluorophores
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DOI：
10.1038/nbt0497-373
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发表时间：
1997-04-01
影响因子：
46.9
通讯作者：
Ulfhake, B
中科院分区：
文献类型：
--
作者：
Brismar, H;Ulfhake, B
为了在离散的组织域中通过其寿命来分辨多个荧光团，标记强度必须足够强，并且标记之间的强度差不能太大，所有荧光团的衰落速率应该相似，并且荧光团的寿命应该足够离散。我们可以很容易地区分花青素-3.18 （Cy-3）、丽丝明罗丹明（LRSC）和德克萨斯红，当它们在组织谱中没有共定位时。Cy-3与LRSC、Cy3与Texas Red的共定位也可以区分，而LRSC与Texas Red的结合则比较困难。我们在共聚焦显微镜中使用荧光寿命记录来检测神经元中的不同神经肽。我们证明生长抑素和丙氨酸在脊髓背角的轴突中是共定位的。
In order to resolve multiple fluorophores by their lifetimes in discrete tissue domains, the labeling intensity must be sufficiently strong and the intensity-difference between the labels must not be too large, the rate of fading should be similar for all fluorophores, and the lifetimes of the fluorophores should be sufficiently discrete. We could readily distinguish Cyanine-3.18 (Cy-3), Lissamine Rhodamine (LRSC), and Texas Red when they were not colocalized in tissue profiles. Colocalization of Cy-3 and LRSC, as well as Cy3 and Texas Red, could also be distinguished, while the combination of LRSC and Texas Red was more difficult. We have used fluorescence lifetime recordings in confocal microscopy to detect different neuropeptides in neurons. We demonstrate that somatostatin and galanin are colocalized in axon profiles of the spinal cord dorsal horn.