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一种用于微阵列制造的高度并行纳升分配器。

A highly parallel nanoliter dispenser for microarray fabrication.

作者信息

Gutmann Oliver, Kuehlewein Ruben, Reinbold Stefanie, Niekrawietz Remigius, Steinert Chris P, de Heij Bas, Zengerle Roland, Daub Martina

机构信息

IMTEK-University of Freiburg, Laboratory for MEMS Applications, Georges-Koehler-Allee 103, D-79110 Freiburg, Germany.

出版信息

Biomed Microdevices. 2004 Jun;6(2):131-7. doi: 10.1023/b:bmmd.0000031750.37323.dd.

Abstract

We report about the correlation between satellite free droplet release and liquid viscosity in a highly parallel, pressure driven nanoliter dispenser. In extensive studies, we found that for liquids of different viscosities the duration of the pressure pulse is the predominant effect compared to pressure amplitude. This result is of essential importance when actuation parameters have to be adopted for different media like oligonucleotide, DNA or protein solutions as it is the case for the non-contact high throughput fabrication of microarrays (Ducree et al., 2000). Experiments with oligonucleotides as well as with different proteins showed ascertained carry-over and cross-contamination free printing of DNA and protein microarrays. With it a prime critical point of microarray production is solved, leading to high quality whilst high throughput microarray fabrication. For oligonucleotides printing, we found CVs to be better than 1% within one single dispensing channel and 1.5% within all 24 channels of a 24 channel printhead for each used printing buffer. By optimizing the protein printing buffer the CVs for protein printing were reduced to about 1% within all 24 channels. As a serious practical application test oligonucleotides microarrays were produced using our nanoliter dispenser system. With it a full DNA hybridization experiment was performed. Clear positive signals one hand and no signals in the negative controls on the other hand showed that our system is suited for microarray production.

摘要

我们报告了在高度平行、压力驱动的纳升分配器中,无卫星液滴释放与液体粘度之间的相关性。在广泛的研究中,我们发现,对于不同粘度的液体,与压力幅度相比,压力脉冲的持续时间是主要影响因素。当必须针对不同介质(如寡核苷酸、DNA或蛋白质溶液)采用驱动参数时,这一结果至关重要,就像微阵列的非接触式高通量制造那样(Ducree等人,2000年)。使用寡核苷酸以及不同蛋白质进行的实验表明,DNA和蛋白质微阵列的打印能够确保无残留和无交叉污染。由此解决了微阵列生产的一个主要关键点,从而实现了高质量且高通量的微阵列制造。对于寡核苷酸打印,我们发现,在每个使用的打印缓冲液中,单个分配通道内的变异系数(CV)优于1%,在24通道打印头的所有24个通道内为1.5%。通过优化蛋白质打印缓冲液,蛋白质打印在所有24个通道内的CV降低到了约1%。作为一项严格的实际应用测试,我们使用纳升分配器系统生产了寡核苷酸微阵列。据此进行了完整的DNA杂交实验。一方面有清晰的阳性信号,另一方面阴性对照中无信号,这表明我们的系统适用于微阵列生产。

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