Nonlinear optical potentiometric dyes optimized for imaging with 1064-nm light.
Nonlinear optical potentiometric dyes optimized for imaging with 1064-nm light.
复制标题
非线性光学电位染料针对 1064 nm 光成像进行了优化。
DOI：
10.1117/1.2772276
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发表时间：
2007
影响因子：
3.5
通讯作者：
Loew,LeslieM
中科院分区：
文献类型：
--
作者：
Teisseyre,ThomasZ;Millard,AndrewC;Yan,Ping;Wuskell,JosephP;Wei,Mei-de;Lewis,Aaron;Loew,LeslieM
非线性光学现象，如双光子荧光(2PF)和二次谐波产生(SHG)，与电压敏感染料相结合，可以获得可兴奋细胞和组织中电活动的高分辨率时空图。1064 nm光纤激光技术的发展简化了高强度、长波长、飞秒脉冲的产生，能够穿透组织深处。要将这两个进步结合起来，需要设计和合成新的染料，这些染料针对更长的波长进行优化，并对膜电位变化产生快速和灵敏的反应。在这项工作中，我们系统地筛选了一系列具有不同发色团和侧链的新染料，这些染料将它们固定在细胞膜上。我们发现了几种可能用于活体测量细胞电活动的染料，因为它们对膜电位的反应迅速而灵敏。其中一些染料表现出最佳的SHG活性；另一些则显示出最佳的2PF活性。这种受监管的染料筛选方法也使我们能够深入了解SHG和2PF背后的分子机制。特别是，这两种非线性光学模式的灵敏度和动力学模式的不同表明，它们的电压灵敏度源于不同的机制。
Nonlinear optical phenomena, such as two-photon fluorescence (2PF) and second harmonic generation (SHG), in combination with voltage sensitive dyes, can be used to acquire high-resolution spatio temporal maps of electrical activity in excitable cells and tissue. Developments in 1064-nm fiber laser technology have simplified the generation of high-intensity, long-wavelength, femtosecond light pulses, capable of penetrating deep into tissue.To merge these two advances requires the design and synthesis of new dyes that are optimized for longer wavelengths and that produce fast and sensitive responses to membrane potential changes. In this work, we have systematically screened a series of new dyes with varying chromophores and sidechains that anchor them in cell membranes. We discovered several dyes that could potentially be used forin vivomeasurements of cellular electrical activity because of their rapid and sensitive responses to membrane potential. Some of these dyes show optimal activity for SHG; others for 2PF. This regulated approach to dye screening also allows significant insight into the molecular mechanisms behind both SHG and 2PF. In particular, the differing patterns of sensitivity and kinetics for these two nonlinear optical modalities indicate that their voltage sensitivity originates from differing mechanisms.