Steuwe Christian, Bové Hannelore, Clerinx Jan, vandeVen Martin, Fron Eduard, Nawrot Tim, Ameloot Marcel, Roeffaers Maarten
Centre for Surface Chemistry and Catalysis, KU Leuven, Leuven, Belgium.
Biomedical Research Institute, Hasselt University, Diepenbeek, Belgium.
J Biophotonics. 2018 May;11(5):e201700233. doi: 10.1002/jbio.201700233. Epub 2018 Jan 30.
Carbonaceous particle exposure and air pollution in general lead to a multitude of adverse human health effects and pose multiple challenges in terms of exposure, risk and safety assessment. Highly desirable for fast screening are label-free approaches for detecting these particle types in biological or medical context. We report a powerful approach for detecting carbonaceous particles using photothermal pump-probe microscopy, which directly probes their strong light absorption. The principle and reliability of this approach is demonstrated by examining 4 different carbon black (CB) species modeling soot with diameters ranging from 13 to 500 nm. Our results show that the proposed approach is applicable to a large number of CB types as well as black carbon. As the particles show a strong absorption over a wide spectral range as compared to other absorbing species, we can image CB particles almost background free. Our pump-probe approach allows label-free optical detection and unambiguous localization of CB particles in (bio)fluids and 3D cellular environments. In combination with fluorescence microscopy, this method allows for simultaneous colocalization of CB with different cellular components using fluorophores as shown here for human lung fibroblasts. We further demonstrate the versatility of pump-probe detection in a flow cell.
一般来说，碳质颗粒暴露和空气污染会导致多种不良的人类健康影响，并在暴露、风险和安全评估方面带来多重挑战。在生物或医学背景下，用于检测这些颗粒类型的无标记方法对于快速筛查非常理想。我们报告了一种使用光热泵浦-探测显微镜检测碳质颗粒的强大方法，该方法直接探测它们的强光吸收。通过检查4种不同的炭黑（CB）物种（模拟直径范围为13至500nm的烟灰），证明了该方法的原理和可靠性。我们的结果表明，所提出的方法适用于大量的CB类型以及黑碳。由于与其他吸收物种相比，这些颗粒在很宽的光谱范围内都有很强的吸收，我们可以在几乎无背景的情况下对CB颗粒进行成像。我们的泵浦-探测方法允许在（生物）流体和3D细胞环境中对CB颗粒进行无标记光学检测和明确的定位。结合荧光显微镜，该方法允许使用荧光团将CB与不同的细胞成分同时进行共定位，如此处针对人肺成纤维细胞所示。我们进一步证明了泵浦-探测检测在流动池中应用的多功能性。
