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On-chip ultrasonic sample preparation for cell based assays

机译：片上超声波样品制备用于基于细胞的测定

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We demonstrate an acoustophoresis method for size-based separation, isolation, up-concentration and trapping of cells that can be used for on-chip sample preparation combined with high resolution imaging for cell-based assays. The method combines three frequency-specific acoustophoresis functions in a sequence by actuating three separate channel zones simultaneously: zones for pre-alignment, size-based separation, and trapping. We characterize the mutual interference between the acoustic radiation forces between the different zones by measuring the spatial distribution of the acoustic energy density during different schemes of ultrasonic actuation, and use this information for optimizing the driving frequencies and voltages of the three utilized ultrasonic transducers attached to the chip, and the flow rates of the pumps. By the use of hydrodynamic defocusing of the pre-aligned cells in the separation zone, a cell population from a complex sample can be isolated and trapped with very high purity, followed by dynamic fluorescence analysis. We exemplify the method's potential by isolating A549 lung cancer cells from red blood cells with 100% purity, 92% separation efficiency, and 93% trapping efficiency resulting in a 130× up-concentration factor during 15 minutes of continuous sample processing through the chip. Furthermore, we demonstrate an on-chip fluorescence assay of the isolated cancer cells by monitoring the dynamic uptake and release of a fluorescence probe in individual trapped cells. The ability to combine isolation of individual cells from a complex sample with high-resolution image analysis holds great promise for applications in cellular and molecular diagnostics.
机译：我们证明了一种用于尺寸的分离，分离，升高和诱捕的辐射磷酸酯方法，其可用于片上样品制剂与高分辨率成像组合用于基于细胞的测定。该方法通过同时致动三个单独的频道区域：用于预取向，基于尺寸的分离和捕获的区域，将三种频率特异性声渗函数函数组合在序列中。我们通过测量超声波致动的不同方案期间声能密度的空间分布来表征不同区域之间的声学​​辐射力之间的相互干扰，并使用该信息优化附着的三个利用超声换能器的驱动频率和电压芯片和泵的流速。通过使用分离区中预先排列的细胞的流体动力学散核，可以用非常高的纯度分离来自复合样品的细胞群，然后具有动态荧光分析。我们通过将A549肺癌细胞与100％纯度，92％分离效率和93％的诱捕效率分离出A549肺癌细胞来举例说明该方法的电位，导致通过芯片的连续样品加工15分钟的130×上升因子。此外，我们通过监测单个被捕获的细胞中的荧光探针的动态吸收和释放来证明分离的癌细胞的片上荧光测定。将单个细胞与高分辨率图像分析分离的单个细胞分离的能力对细胞和分子诊断中的应用具有很大的希望。

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