平行DLD微流体用于叶绿体分离和分选

IF 5.4 2区 工程技术 Q1 BIOCHEMICAL RESEARCH METHODS
Lab on a Chip Pub Date : 2025-06-09 DOI:10.1039/d5lc00348b
Oriana Gerallin Chavez-Pineda, Pablo E. Guevara-Pantoja, Victor Marin, Gabriel Arturo Caballero-Robledo, Luis David Patino-Lopez, Daniel A May-Arrioja, Clelia De-la-Peña, Jose L. Garcia-Cordero
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引用次数: 0

摘要

叶绿体是植物细胞特有的细胞器,对光合作用和各种其他代谢过程至关重要,包括氨基酸、脂质和激素的生物合成。除了它们的传统功能,叶绿体已经成为生物技术中有前途的目标,特别是在治疗应用和生物燃料生产方面。然而,由于传统方法的局限性,它们的隔离在技术上仍然具有挑战性,通常需要复杂的协议、专门的设备和训练有素的人员。在这里,我们提出了一个基于微流体的平台,可以使用确定性横向位移(DLD)实现基于尺寸的叶绿体分离。我们的器件集成了四个并行DLD阵列,每个阵列都有不同的临界直径(CD)。该配置可实现带通滤波,并允许在单个设备内同时分离各种大小的叶绿体。与传统技术相比,所有阵列的共享入口和均匀流动条件提高了再现性。传统的蔗糖密度梯度缺乏精确的基于尺寸的分离,与此不同,我们的系统对3至8µm的叶绿体的分离效率为50-85%,回收的组分纯度为17-66%。该平台为叶绿体分离提供了快速、自动化和可扩展的解决方案,在植物研究、生物技术和合成生物学方面具有重要的潜在应用。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Parallel DLD Microfluidics for Chloroplast Isolation and Sorting
Chloroplasts are characteristic organelles of plant cells, essential for photosynthesis and various other metabolic processes, including amino acid, lipid, and hormone biosynthesis. Beyond their classical functions, chloroplasts have emerged as promising targets in biotechnology, particularly in therapeutic applications and biofuel production. However, their isolation remains technically challenging due to the limitations of conventional methods, which typically require complex protocols, specialized equipment, and trained personnel. Here, we present a microfluidic-based platform that enables size-based chloroplast separation using deterministic lateral displacement (DLD). Our device integrates four parallel DLD arrays, each with a distinct critical diameter (CD). This configuration enables bandpass filtering and allows the simultaneous isolation of chloroplasts of various sizes within a single device. Shared inlets and uniform flow conditions across all arrays enhance reproducibility compared to conventional techniques. Unlike traditional sucrose density gradients, which lack precise size-based separation, our system achieves separation efficiencies of 50-85% for chloroplasts ranging from 3 to 8 µm, with recovered fractions having purities of 17-66%. This platform provides a rapid, automated, and scalable solution for chloroplast isolation, with significant potential applications in plant research, biotechnology, and synthetic biology.
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来源期刊
Lab on a Chip
Lab on a Chip 工程技术-化学综合
CiteScore
11.10
自引率
8.20%
发文量
434
审稿时长
2.6 months
期刊介绍: Lab on a Chip is the premiere journal that publishes cutting-edge research in the field of miniaturization. By their very nature, microfluidic/nanofluidic/miniaturized systems are at the intersection of disciplines, spanning fundamental research to high-end application, which is reflected by the broad readership of the journal. Lab on a Chip publishes two types of papers on original research: full-length research papers and communications. Papers should demonstrate innovations, which can come from technical advancements or applications addressing pressing needs in globally important areas. The journal also publishes Comments, Reviews, and Perspectives.
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