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基于微流控技术的单细胞分析揭示了锥虫中药物依赖性的运动变化。

Microfluidics-based single cell analysis reveals drug-dependent motility changes in trypanosomes.

作者信息

Hochstetter Axel, Stellamanns Eric, Deshpande Siddharth, Uppaluri Sravanti, Engstler Markus, Pfohl Thomas

机构信息

Department of Chemistry, University of Basel, 4056 Basel, Switzerland.

出版信息

Lab Chip. 2015 Apr 21;15(8):1961-8. doi: 10.1039/c5lc00124b.

Abstract

We present a single cell viability assay, based on chemical gradient microfluidics in combination with optical micromanipulation. Here, we used this combination to in situ monitor the effects of drugs and chemicals on the motility of the flagellated unicellular parasite Trypanosoma brucei; specifically, the local cell velocity and the mean squared displacement (MSD) of the cell trajectories. With our method, we are able to record in situ cell fixation by glutaraldehyde, and to quantify the critical concentration of 2-deoxy-d-glucose required to completely paralyze trypanosomes. In addition, we detected and quantified the impact on cell propulsion and energy generation at much lower 2-deoxy-d-glucose concentrations. Our microfluidics-based approach advances fast cell-based drug testing in a way that allows us to distinguish cytocidal from cytostatic drug effects, screen effective dosages, and investigate the impact on cell motility of drugs and chemicals. Using suramin, we could reveal the impact of the widely used drug on trypanosomes: suramin lowers trypanosome motility and induces cell-lysis after endocytosis.

摘要

我们展示了一种基于化学梯度微流控技术与光学微操纵相结合的单细胞活力测定方法。在此,我们利用这种组合原位监测药物和化学物质对有鞭毛的单细胞寄生虫布氏锥虫运动性的影响;具体而言,是细胞的局部速度和细胞轨迹的均方位移(MSD)。通过我们的方法,我们能够记录戊二醛原位固定细胞的情况,并量化完全麻痹锥虫所需的2-脱氧-D-葡萄糖的临界浓度。此外,我们在低得多的2-脱氧-D-葡萄糖浓度下检测并量化了对细胞推进和能量产生的影响。我们基于微流控的方法推动了快速的基于细胞的药物测试,使我们能够区分细胞毒性和细胞抑制性药物作用,筛选有效剂量,并研究药物和化学物质对细胞运动性的影响。使用苏拉明,我们能够揭示这种广泛使用的药物对锥虫的影响:苏拉明降低锥虫的运动性,并在内吞作用后诱导细胞裂解。

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