Simultaneous Photographing of Oxygen and pH In Vivo Using Sensor Films
Simultaneous Photographing of Oxygen and pH In Vivo Using Sensor Films
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DOI：
10.1002/anie.201104530
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发表时间：
2011-01-01
影响因子：
16.6
通讯作者：
Wolfbeis, Otto S.
中科院分区：
文献类型：
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作者：
Meier, Robert J.;Schreml, Stephan;Wolfbeis, Otto S.
体内化学分析物（例如 CO2、[1] H2O2、[2] 氧、[3] Ca2+、[4] 或 pH [5]）的光学成像是一个快速发展的领域，因为这些物质可以在整个区域而不是在单个测量点进行观察和量化。这一特征在生物医学科学中尤其重要，因为即使在很小的区域（例如细胞）中也可以发现此类参数的巨大变化，并且采样通常不可能或会改变分析物的实际浓度（例如在 O2 的情况下）。此类物质的分布通常使用基于发光的读出技术来可视化，[6]，这些技术可以基于强度、[7] 荧光寿命 (FLIM)、[1, 8] 或两个波长信号（比率测定法）[9] 的测量。为了实现定量成像并消除荧光探针浓度和分布变化造成的影响，必须进行参考。（有关参考方法的介绍，请参见支持信息，注1。）这里我们介绍第一种通过传统数码相机仅获取一张图像同时成像两个参数的方法。放置在相机前面的 LED 阵列用于光激发。这里的相机还充当基本光谱仪，用于具有一个激发波长和多个（三波长）读数的比率成像。采用带有固定指示探针的传感器薄膜，而不是溶解的探针。
Optical imaging of chemical analytes such as CO2,[1] H2O2,[2] oxygen,[3] Ca2+,[4] or pH [5] invivo is a fast-growing field because these species can be observed and quantified over whole areas rather than at a single point of measurement. This feature is particularly important in biomedical sciences where large variations of such parameters can be found in even small areas (such as cells), and where sampling usually is impossible or would alter the actual concentration of the analyte (for example in the case of O2). The distribution of such species is most often visualized using luminescence-based readout techniques,[6] and these can be based on measurement of intensity,[7] fluorescence lifetime (FLIM),[1, 8] or of signals at two wavelengths (ratiometry),[9] for example. Referencing is mandatory to enable quantitative imaging and to eliminate effects caused by variations in the concentration and distribution of fluorescent probes.(For an introduction into methods for referencing, see the Supporting Information, Note 1.)Herein we introduce the first method for simultaneous imaging of two parameters with only one image acquired by a conventional digital camera. An LED array placed in front of the camera is used for photoexcitation. The camera here also acts as a rudimental spectrometer for ratiometric imaging with one excitation wavelength and multiple (three-wavelength) readout. A sensor film with immobilized indicator probes is employed rather than dissolved probes which have