CARS microscopy lights up lipids in living cells
CARS microscopy lights up lipids in living cells
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发表时间：
2004-08
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通讯作者：
Xiaolin Nan;Wei Yang;X. Xie
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作者：
Xiaolin Nan;Wei Yang;X. Xie
这些技术，在未染色的样品中具有化学选择性成像。CARS显微镜是一种非线性成像技术，它根据化学物种的振动特征生成它们的图像。在过去的五年里，对该方法的对比机制和可用的仪器设备的理解都取得了进展。1传统的激光扫描显微镜可以在不改变样品的情况下提供高灵敏度和选择性的对比度，这是生物学家希望的成像技术。荧光显微镜具有很高的灵敏度，但染料可以改变样品。相位对比显微镜和差分干涉对比显微镜避免了这个问题，但代价是化学专一性。相干反斯托克斯拉曼散射(CARS)显微镜适合CARS显微镜照亮活细胞中的脂类，以1到2 mW的良性激发功率和视频的图像采集速率进行这些成像实验。这项技术为许多新兴和令人兴奋的生物学应用提供了曙光。在它揭示的活细胞中的化学物种中，2种脂类提供了最好的对比，为加强肥胖和动脉粥样硬化等脂类相关疾病的生物医学研究提供了巨大的潜力。CARS显微镜依靠拉曼效应。在自发拉曼过程中，分子散射光子，用与分子振动模式相对应的能量量子来修改光子能量。因此，自发拉曼活细胞成像图。1.在拉曼散射过程中，入射光根据分子的振动频率(Ω)改变频率。红移和蓝移分量分别称为斯托克斯线和反斯托克斯线。在自发拉曼(A)中，分子的热驱动和随机相振动导致在各个方向上的低效散射。在相干反斯托克斯拉曼散射(B)中，频率为ωp和ωS的两束激发光形成频率为ωpϪωS的拍频光场，当ωpϪωS匹配Ω时，分子振动发生同相高效，产生强烈的方向性信号。
these techniques, imaging with chemical selectivity in unstained samples. CARS microscopy is a nonlinear imaging technique that produces images of chemical species based on their vibra-tional signatures. The past five years have seen advances in both the understanding of the method's contrast mechanism and the available instrumentation. 1 Conventional laser scanning microscopes can rou-B iologists desire imaging techniques that provide contrast with high sensitivity and selectivity but without altering the sample. Fluorescence mi-croscopy has high sensitivity, but dyes can alter the sample. Phase-contrast and differential-interference-contrast microscopes avoid this problem at the cost of chemical specificity. Coherent anti-Stokes Raman scattering (CARS) microscopy fits CARS Microscopy Lights Up Lipids in Living Cells tinely carry out these imaging experiments with benign excitation powers of 1 to 2 mW and image acquisition rates up to that of video. The technique is shedding light on many emerging and exciting biological applications. Among the chemical species that it has revealed in living cells, 2 lipids provide the best contrast, offering great potential to augment biomedical research in lipid-related diseases such as obesity and atherosclerosis. CARS microscopy relies on the Raman effect. In the spontaneous Raman process, molecules scatter photons, modifying the photon energy with energy quanta that corresponds to the molecules' vibrational modes. Hence, spontaneous Raman LIVE-CELL IMAGING Figure. 1. In Raman scattering processes, incoming light changes frequency according to a vibrational frequency (Ω) of the molecules. Red-and blue-shifted components are termed Stokes and anti-Stokes lines, respectively. In spontaneous Raman (A), thermally driven and random-phased molecular vibrations cause inefficient scattering in all directions. In coherent anti-Stokes Raman scattering (B), two excitation beams at frequencies ω p and ω s form a beating field with frequency ω p Ϫ ω s. When ω p Ϫ ω s matches Ω, the molecular vibrations occur in-phase and efficiently, resulting in a strong directional signal.