Detection of drug-induced cellular changes using confocal Raman spectroscopy on patterned single-cell biosensors.
Detection of drug-induced cellular changes using confocal Raman spectroscopy on patterned single-cell biosensors.
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DOI：
10.1039/b900420c
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发表时间：
2009-07
期刊：
影响因子：
--
通讯作者：
Zhang M
中科院分区：
文献类型：
--
作者：
Buckmaster R;Asphahani F;Thein M;Xu J;Zhang M
我们报告了一个基于细胞的生物传感器的应用，利用图案化的单细胞阵列结合共焦拉曼光谱观察时间依赖性药物反应的单个细胞在真实的时间。图案化的单细胞平台使得能够容易地定位单个细胞并且连续地可寻址，以用于以非破坏性的定量方式对生化组成变化进行拉曼光谱表征，从而保留离散的细胞行为和细胞间变化。在这项研究中，人髓母细胞瘤（DAOY）细胞暴露于常见的化疗剂依托泊苷，并从图案化的细胞的拉曼光谱记录超过48小时。结果发现，药物暴露48小时后，监测的87.5%的细胞显示DNA和蛋白质相关峰急剧下降，对应于细胞死亡。其余12.5%的细胞显示关键拉曼生物标志物几乎没有减少，表明它们具有耐药性。此外，通过流式细胞术证实，图案化细胞群显示出与融合细胞培养物非常相似的对依托泊苷的反应。最后，用TUNEL法测定依托泊苷暴露后由于DNA片段化引起的细胞凋亡。结果与拉曼光谱分析结果吻合较好。这种组合的生物传感器-拉曼平台提供了一种快速，简单的方法来评估细胞对化学和生物制剂的反应，具有高通量，并可用于各种生物医学应用，如药物发现，毒素测试和生物威胁检测。
We report on a cell-based biosensor application that utilizes patterned single-cell arrays combined with confocal Raman spectroscopy to observe the time-dependent drug response of individual cells in real time. The patterned single-cell platform enables individual cells to be easily located and continuously addressable for Raman spectroscopy characterization of biochemical compositional changes in a non-destructive, quantitative manner so that discrete cellular behavior and cell-to-cell variations are preserved. In this study, human medulloblastoma (DAOY) cells were exposed to the common chemotherapeutic agent etoposide, and Raman spectra from patterned cells were recorded over 48 hours. It was found that 87.5% of cells monitored exhibited a sharp decrease in DNA and protein associated peaks 48 hours after drug exposure, corresponding to cell death. The remaining 12.5% of cells showed little to no reduction in key Raman biomarkers, indicating their drug resistance. Furthermore, the patterned cell population showed a very similar response to etoposide as confluent cell cultures as confirmed by flow cytometry. Finally, patterned cells were assessed with TUNEL assay for apoptosis due to DNA fragmentation after etoposide exposure. The results agree well with those from the Raman spectroscopy analysis. This combined biosensor-Raman platform provides a quick, simple way to assess cell responses to chemical and biological agents with high throughput and can be potentially used for a wide variety of biomedical applications such as pharmaceutical drug discovery, toxin tests, and biothreat detection.
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影响因子：
3.9
作者：
Veiseh, M;Zareie, MH;Zhang, MQ
通讯作者：
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