Pikov Victor, Siegel Peter H
Neural Engineering Program of the Huntington Medical Research Institutes.
IEEE Eng Med Biol Mag. 2010 Jan-Feb;29(1):63-71. doi: 10.1109/MEMB.2009.935468.
A simple setup is demonstrated for remote temperature monitoring of water, water-based media, and cells on a microscopic scale. The technique relies on recording changes in the shape of a stretching band of the hydroxyl group in liquid water at 3,100-3,700 cm(-1). Rather than direct measurements in the near-infrared (IR), a simple Raman spectrometer setup is realized. The measured Raman shifts are observed at near optical wavelengths using an inverted microscope with standard objectives in contrast to costly near-IR elements. This allows for simultaneous visible inspection through the same optical path. An inexpensive 671-nm diode pump laser (< 100 mW), standard dichroic and lowpass filters, and a commercial 600-1,000 nm spectrometer complete the instrument. Temperature changes of 1 degrees C are readily distinguished over a range consistent with cellular processes (25-45 degrees C) using integration times below 10 s. Greatly improved sensitivity was obtained by an automated two-peak fitting procedure. When combined with an optical camera, the instrument can be used to monitor changes in cell behavior as a function of temperature without the need for invasive probing. The instrument is very simple to realize, inexpensive compared with traditional Raman spectrometers and IR microscopes, and applicable to a wide range of problems in microthermometry of biological systems. In a first application of its kind, the instrument was used to successfully determine the temperature rise of a cluster of H1299 derived human lung cells adhered to polystyrene and immersed in phosphate-buffered saline (PBS) under exposure of RF millimeter wave radiation (60 GHz, 1.3, 2.6, and 5.2 mW/mm2).
展示了一种简单的装置，用于在微观尺度上对水、水基介质和细胞进行远程温度监测。该技术依赖于记录液态水中羟基伸缩带在3100 - 3700 cm(-1)处形状的变化。无需在近红外（IR）进行直接测量，而是实现了一种简单的拉曼光谱仪装置。与昂贵的近红外元件不同，使用带有标准物镜的倒置显微镜在近光学波长下观察测量的拉曼位移。这允许通过相同的光路同时进行可见检查。一个廉价的671 - nm二极管泵浦激光器（<100 mW）、标准二向色镜和低通滤波器以及一台商用600 - 1000 nm光谱仪构成了该仪器。使用低于10 s的积分时间，在与细胞过程一致的温度范围（25 - 45℃）内，很容易分辨出1℃的温度变化。通过自动双峰拟合程序获得了大大提高的灵敏度。当与光学相机结合使用时，该仪器可用于监测细胞行为随温度的变化，而无需进行侵入性探测。该仪器实现起来非常简单，与传统拉曼光谱仪和红外显微镜相比成本较低，适用于生物系统微热测量中的广泛问题。在同类的首次应用中，该仪器成功用于确定附着在聚苯乙烯上并浸没在磷酸盐缓冲盐水（PBS）中的源自H1299的人肺细胞簇在射频毫米波辐射（60 GHz，1.3、2.6和5.2 mW/mm2）照射下的温度升高。
