An Open-Source Plate Reader
An Open-Source Plate Reader
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DOI：
10.1021/acs.biochem.8b00952
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发表时间：
2019-02-12
期刊：
影响因子：
2.9
通讯作者：
Chow, Brian Y.
中科院分区：
文献类型：
--
作者：
Szymula, Karol P.;Magaraci, Michael S.;Chow, Brian Y.
微板阅读器是实验生物学和生物工程中的基础仪器，可以实现多路分光光度测量。为了增强他们的可及性，我们在这里报告了一个开源微板阅读器的设计、构造、验证和基准测试。该系统具有全光谱吸收和荧光发射检测、原位光遗传刺激以及自动分析协议的独立触摸屏编程。整个系统的成本不到3500美元，只是商业平板阅读器成本的一小部分，可以在低至10 nm的浓度下检测常见染料的荧光。在合成生物学、光遗传学和光传感生物学的背景下，通过稳态测量配体诱导的报告基因在细菌群体感应模型中的表达，以及通过黄素光循环动力学测量用于光遗传转录激活的LOV(光氧电压)结构域光感受器，证明了功能能力。提供了使用定制的基于Python的API(应用程序接口)组装设备并使其自动化的完全详细的指南。这项工作为不断增长的社区范围内以生物为中心的开源硬件基础设施贡献了一项关键技术，这些硬件的创建得益于快速原型能力和低成本的电子、光电子学和微型计算机。
Microplate readers are foundational instruments in experimental biology and bioengineering that enable multiplexed spectrophotometric measurements. To enhance their accessibility, we here report the design, construction, validation, and benchmarking of an open-source microplate reader. The system features full-spectrum absorbance and fluorescence emission detection, in situ optogenetic stimulation, and stand-alone touch screen programming of automated assay protocols. The total system costs less than $3500, a fraction of the cost of commercial plate readers, and can detect the fluorescence of common dyes at concentrations as low as, similar to 10 nM. Functional capabilities were demonstrated in the context of synthetic biology, optogenetics, and photosensory biology: by steady-state measurements of ligand-induced reporter gene expression in a model of bacterial quorum sensing and by flavin photocycling kinetic measurements of a LOV (light-oxygen-voltage) domain photoreceptor used for optogenetic transcriptional activation. Fully detailed guides for assembling the device and automating it using the custom Python-based API (Application Program Interface) are provided. This work contributes a key technology to the growing community-wide infrastructure of open-source biology-focused hardware, whose creation is facilitated by rapid prototyping capabilities and low-cost electronics, optoelectronics, and microcomputers.