Long-term two-photon fluorescence imaging of mammalian embryos without compromising viability
Long-term two-photon fluorescence imaging of mammalian embryos without compromising viability
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DOI：
10.1038/11698
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发表时间：
1999-08-01
影响因子：
46.9
通讯作者：
Bavister, BD
中科院分区：
文献类型：
--
作者：
Squirrell, JM;Wokosin, DL;Bavister, BD
活哺乳动物细胞的荧光成像的主要挑战是在长时间暴露于激发照明之后保持活力。我们使用双光子显微镜（1，047 nm）在24小时内频繁监测仓鼠胚胎中线粒体分布的动态，同时保持胚泡甚至胎儿的发育能力。相比之下，共聚焦成像仅8小时抑制发展，即使没有荧光团的激发，光诱导产生的H2O2可能占，在某种程度上，这种抑制，因此，双光子显微镜，但不共聚焦显微镜，已允许长期荧光观察的动态三维细胞结构在高度感光的标本，如哺乳动物胚胎。
A major challenge for fluorescence imaging of living mammalian cells is maintaining viability following prolonged exposure to excitation illumination. We have monitored the dynamics of mitochondrial distribution in hamster embryos at frequent intervals over 24 h using two-photon microscopy (1,047 nm) while maintaining blastocyst, and even fetal, developmental competence. In contrast, confocal imaging for only 8 h inhibits development, even without fluorophore excitation, Photo-induced production of H2O2 may account, in part, for this inhibition, Thus, two-photon microscopy, but not confocal microscopy, has permitted long-term fluorescence observations of the dynamics of three-dimensional cytoarchitecture in highly photosensitive specimens such as mammalian embryos.