Lucas Pierre, Le Coq David, Juncker Christophe, Collier Jayne, Boesewetter Dianne E, Boussard-Plédel Catherine, Bureau Bruno, Riley Mark R
Department of Material Science and Engineering, University of Arizona, Tucson, AZ 85721, USA.
Appl Spectrosc. 2005 Jan;59(1):1-9. doi: 10.1366/0003702052940387.
Biochemical changes in living cells are detected using a fiber probe system composed of a single chalcogenide fiber acting as both the sensor and transmission line for infrared optical signals. The signal is collected via evanescent wave absorption along the tapered sensing zone of the fiber. We spectroscopically monitored the effects of the surfactant Triton X-100, which serves as a toxic agent simulant on a transformed human lung carcinoma type II epithelial cell line (A549). We observe spectral changes between 2800-3000 cm(-1) in four absorptions bands, which are assigned to hydrocarbon vibrations of methylene and methyl groups in membrane lipids. Comparison of fiber and transmission spectra shows that the present technique allows one to locally probe the cell plasma membrane in the lipid spectral region. These optical responses are correlated with cellular metabolic activity measurements and LDH (lactate dehydrogenase) release assays that indicate a loss of cellular function and membrane integrity as would be expected in response to the membrane solubilizing Triton. The spectroscopic technique shows a significantly greater detection resolution in time and concentration.
利用一种由单一硫族化物光纤组成的光纤探针系统来检测活细胞中的生化变化,该光纤同时充当红外光信号的传感器和传输线。信号通过沿光纤锥形传感区的倏逝波吸收来收集。我们用光谱法监测了表面活性剂吐温X-100(它用作有毒剂模拟物)对转化的人II型肺癌上皮细胞系(A549)的影响。我们在2800 - 3000 cm(-1)之间的四个吸收带中观察到光谱变化,这些吸收带归因于膜脂中亚甲基和甲基的烃振动。光纤光谱与透射光谱的比较表明,本技术能够在脂质光谱区域局部探测细胞质膜。这些光学响应与细胞代谢活性测量以及乳酸脱氢酶(LDH)释放测定相关,这些测量和测定表明细胞功能丧失和膜完整性受损,这正如预期的对膜增溶的吐温的反应。该光谱技术在时间和浓度上显示出显著更高的检测分辨率。