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首页> 外文会议>Photons Plus Ultrasound: Imaging and Sensing 2007; Progress in Biomedical Optics and Imaging; vol.8 no.14; Proceedings of SPIE-The International Society for Optical Engineering; vol.6437 >Imaging living cells with a combined high-resolution multi-photonacoustic microscope
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Imaging living cells with a combined high-resolution multi-photonacoustic microscope

机译：组合高分辨率多光声显微镜对活细胞成像

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摘要

With increasing demand for in-vivo observation of living cells, microscope techniques that do not need staining become more and more important. In this talk we present a combined multi-photon-acoustic microscope with the possibility to measure synchronously properties addressed by ultrasound and two-photon fluorescence. Ultrasound probes the local mechanical properties of a cell, while the high resolution image of the two-photon fluorescence delivers insight in cell morphology and activity. In the acoustic part of the microscope an ultrasound wave, with a frequency of GHz, is focused by an acoustic sapphire lens and detected by a piezo electric transducer assembled to the lens. The achieved lateral resolution is in the range of 1 μm. Contrast in the images arises mainly from the local absorption of sound in the cells, related to properties, such as mass density, stiffness and viscose damping. Additionally acoustic microscopy can access the cell shape and the state of the cell membrane as it is a intrinsic volume scanning technique.The optical part bases on the emission of fluorescent biomolecules naturally present in cells (e.g. NAD(P)H, protophorphyrin IX, lipofuscin, melanin). The nonlinear effect of two-photon absorption provides a high lateral and axial resolution without the need of confocal detection. In addition, in the near-IR cell damages are drastically reduced in comparison to direct excitation in the visible or UV. Both methods can be considered as minimal invasive, as they relay on intrinsic contrast mechanisms and dispense with the need of staining. First results on living cells are presented and discussed.
机译：随着对活细胞的体内观察的需求增加，不需要染色的显微镜技术变得越来越重要。在本次演讲中，我们介绍了一种组合式多光子声显微镜，可以同时测量超声和双光子荧光所解决的特性。超声探测细胞的局部机械性能，而双光子荧光的高分辨率图像则可洞悉细胞的形态和活性。在显微镜的声学部分，超声蓝宝石透镜聚焦频率为GHz的超声波，并由组装在透镜上的压电换能器检测。达到的横向分辨率在1μm的范围内。图像中的对比度主要来自单元中声音的局部吸收，与诸如质量密度，刚度和粘胶阻尼等特性有关。此外，声波显微镜技术可以检测细胞的形状和细胞膜的状态，因为它是一种固有的体积扫描技术。光学部分基于细胞中天然存在的荧光生物分子（例如NAD（P）H，原卟啉IX，脂褐素）的发射，黑色素）。双光子吸收的非线性效应提供了高的横向和轴向分辨率，而无需共聚焦检测。另外，与在可见光或紫外光中直接激发相比，在近红外中，细胞损伤大大减少。两种方法都可以被认为是微创的，因为它们依靠内在的对比机制并不需要染色。提出并讨论了有关活细胞的初步结果。

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