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开源液体处理仪的技术升级以支持细菌菌落筛选。

Technical upgrade of an open-source liquid handler to support bacterial colony screening.

作者信息

Del Olmo Lianes Irene, Yubero Pablo, Gómez-Luengo Álvaro, Nogales Juan, Espeso David R

机构信息

Department of Systems Biology, Centro Nacional de Biotecnología-Consejo Superior de Investigaciones Científicas, Madrid, Spain.

Interdisciplinary Platform for Sustainable Plastics Towards a Circular Economy-Consejo Superior de Investigaciones Científicas, SusPlast-CSIC, Madrid, Spain.

出版信息

Front Bioeng Biotechnol. 2023 Jun 19;11:1202836. doi: 10.3389/fbioe.2023.1202836. eCollection 2023.

DOI:10.3389/fbioe.2023.1202836
PMID:37404684
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10315574/
Abstract

The optimization of genetically engineered biological constructs is a key step to deliver high-impact biotechnological applications. The use of high-throughput DNA assembly methods allows the construction of enough genotypic variants to successfully cover the target design space. This, however, entails extra workload for researchers during the screening stage of candidate variants. Despite the existence of commercial colony pickers, their high price excludes small research laboratories and budget-adjusted institutions from accessing such extensive screening capability. In this work we present COPICK, a technical solution to automatize colony picking in an open-source liquid handler Opentrons OT-2. COPICK relies on a mounted camera to capture images of regular Petri dishes and detect microbial colonies for automated screening. COPICK's software can then automatically select the best colonies according to different criteria (size, color and fluorescence) and execute a protocol to pick them for further analysis. Benchmark tests performed for and colonies delivers a raw picking performance over pickable colonies of 82% with an accuracy of 73.4% at an estimated rate of 240 colonies/h. These results validate the utility of COPICK, and highlight the importance of ongoing technical improvements in open-source laboratory equipment to support smaller research teams.

摘要

基因工程生物构建体的优化是实现高影响力生物技术应用的关键步骤。高通量DNA组装方法的使用能够构建足够多的基因型变体,从而成功覆盖目标设计空间。然而,这在候选变体的筛选阶段给研究人员带来了额外的工作量。尽管存在商用菌落挑选仪,但因其价格高昂,小型研究实验室和预算有限的机构无法获得这种广泛的筛选能力。在这项工作中,我们展示了COPICK,这是一种用于在开源液体处理仪Opentrons OT-2中实现菌落挑选自动化的技术解决方案。COPICK依靠一个安装的摄像头来捕捉常规培养皿的图像并检测微生物菌落以进行自动筛选。然后,COPICK的软件可以根据不同标准(大小、颜色和荧光)自动选择最佳菌落,并执行一个协议来挑选它们进行进一步分析。对[X]个和[X]个菌落进行的基准测试表明,在估计每小时240个菌落的速度下,可挑选菌落的原始挑选性能为82%,准确率为73.4%。这些结果验证了COPICK的实用性,并突出了开源实验室设备持续技术改进对支持较小研究团队的重要性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2e7b/10315574/56a99a2b374d/fbioe-11-1202836-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2e7b/10315574/7fcde92cc5ca/fbioe-11-1202836-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2e7b/10315574/dfe9141704ed/fbioe-11-1202836-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2e7b/10315574/709d26b64b7b/fbioe-11-1202836-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2e7b/10315574/47c82b9a0f77/fbioe-11-1202836-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2e7b/10315574/c5ecd9ef1537/fbioe-11-1202836-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2e7b/10315574/13bb73e61343/fbioe-11-1202836-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2e7b/10315574/56a99a2b374d/fbioe-11-1202836-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2e7b/10315574/7fcde92cc5ca/fbioe-11-1202836-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2e7b/10315574/dfe9141704ed/fbioe-11-1202836-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2e7b/10315574/709d26b64b7b/fbioe-11-1202836-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2e7b/10315574/47c82b9a0f77/fbioe-11-1202836-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2e7b/10315574/c5ecd9ef1537/fbioe-11-1202836-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2e7b/10315574/13bb73e61343/fbioe-11-1202836-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2e7b/10315574/56a99a2b374d/fbioe-11-1202836-g007.jpg

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