UV–Vis Spectra-Activated Droplet Sorting for Label-Free Chemical Identification and Collection of Droplets,Analytical Chemistry

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UV–Vis Spectra-Activated Droplet Sorting for Label-Free Chemical Identification and Collection of Droplets
Analytical Chemistry ( IF 6.7 ) Pub Date : 2021-09-17 , DOI: 10.1021/acs.analchem.1c02822
Todd A Duncombe _{1,

2} , Aaron Ponti ₁ , Florian P Seebeck _{2,

3} , Petra S Dittrich _{1,

2}

Affiliation

We introduce the UV–vis spectra-activated droplet sorter (UVADS) for high-throughput label-free chemical identification and enzyme screening. In contrast to previous absorbance-based droplet sorters that relied on single-wavelength absorbance in the visible range, our platform collects full UV–vis spectra from 200 to 1050 nm at up to 2100 spectra per second. Our custom-built open-source software application, “SpectraSorter,” enables real-time data processing, analysis, visualization, and selection of droplets for sorting with any set of UV–vis spectral features. An optimized UV–vis detection region extended the absorbance path length for droplets and allowed for the direct protein quantification down to 10 μM of bovine serum albumin at 280 nm. UV–vis spectral data can distinguish a variety of different chemicals or spurious events (such as air bubbles) that are inaccessible at a single wavelength. The platform is used to measure ergothionase enzyme activity from monoclonal microcolonies isolated in droplets. In a label-free manner, we directly measure the ergothioneine substrate to thiourocanic acid product conversion while tracking the microcolony formation. UVADS represents an important new tool for high-throughput label-free in-droplet chemical analysis.

中文翻译：

用于无标记化学物质识别和液滴收集的紫外-可见光谱激活液滴分选

我们介绍了用于高通量无标记化学鉴定和酶筛选的紫外-可见光谱激活液滴分选仪 (UVADS)。与之前依赖于可见光范围内单波长吸光度的基于吸光度的液滴分选机相比，我们的平台以每秒高达 2100 个光谱的速度收集从 200 到 1050 nm 的全紫外可见光谱。我们定制的开源软件应用程序“SpectraSorter”支持实时数据处理、分析、可视化和液滴选择，以使用任何紫外-可见光谱特征进行分选。优化的 UV-vis 检测区域延长了液滴的吸收路径长度，并允许在 280 nm 处将蛋白质直接定量低至 10 μM 的牛血清白蛋白。UV-vis 光谱数据可以区分在单一波长下无法访问的各种不同的化学物质或虚假事件（例如气泡）。该平台用于测量从液滴中分离的单克隆微菌落的麦角硫磷酶活性。我们以无标记的方式直接测量麦角硫因底物到硫脲酸产物的转化，同时跟踪微菌落形成。UVADS 代表了一种重要的新工具，可用于高通量无标记液滴内化学分析。

更新日期：2021-09-28

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