利用细胞外囊泡颗粒计数可视化平台对纳升血浆样品进行疾病诊断

IF 6.7 1区化学 Q1 CHEMISTRY, ANALYTICAL

Analytical Chemistry Pub Date : 2025-02-03 DOI:10.1021/acs.analchem.4c04977

Bing Li, Qunfei Tai, Hailong Yu, Jiaxi Wang, Mingxia Gao, Xiangmin Zhang

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引用次数: 0

摘要

由于细胞外囊泡（EVs）反映了体内病理和生理状态的动态变化，并且在体内具有优异的稳定性，因此它是液体活检应用中无创诊断和预后评估的理想候选者。在此，我们开发了一种单细胞外囊泡荧光可视化和计数平台，该平台由三部分组成：用于抽吸样品的纳流-液操作平台、用于捕获靶向ev的抗体功能化盖板，以及用于ev可视化和计数的单分子荧光标记和成像系统。与之前报道的研究相比，我们开发的平台不仅最大限度地降低了操作复杂性，而且使用最小体积的25 nL等离子体成功检测了单个电动汽车。此外，荧光斑点计数结果表明，开发的平台具有较宽的动态范围，荧光强度与颗粒数之间具有良好的线性相关性，可用于检测标准样品中的CD9+ ev。值得注意的是，我们开发的单个ev捕获和分析平台不仅成功地应用于EpCAM+ ev到CD9+ ev的定量分析，用于乳腺癌（BC）的诊断，还成功地应用于CD14+ ev到CD9+ ev的定量分析，用于2型糖尿病（T2DM）的诊断。总的来说，我们的工作为乳腺癌和2型糖尿病的诊断提供了新的思路，同时由于抗体的可替代性，我们开发了一种通过同时检测痕量样品中Exos亚型的多种生物标志物来区分其他疾病的通用方法。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

Nanoliter Plasma Sample for Disease Diagnosis by Using a Visualization Platform of Counting Extracellular Vesicle Particles

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Nanoliter Plasma Sample for Disease Diagnosis by Using a Visualization Platform of Counting Extracellular Vesicle Particles

Extracellular vesicles (EVs) are an excellent candidate for noninvasive diagnostic and prognostic assessments for liquid biopsy applications owing to reflecting the dynamic changes of pathological and physiological states in the body and exceptional stability in vivo. Herein, we developed a single-extracellular vesicle fluorescence visualization and counting platform containing three parts of the nanoliter-liquid operating platform for aspirating the sample, antibody-functionalized coverslip for capturing targeted EVs, and the single-molecule fluorescence labeling and imaging system for the visualization and counting of EVs. Compared to previously reported studies, our developed platform not only minimized operational complexity but also successfully detected single EVs using a minimal volume of 25 nL of plasma. Moreover, the results of fluorescence spot counts demonstrated that the developed platform exhibits a wide dynamic range and an excellent linear correlation between fluorescence intensity and particle numbers for the detection of CD9⁺ EVs from standard samples. Remarkably, our developed single EVs capture and analysis platform was successfully applied not only in the quantitative analysis of EpCAM⁺ EVs to CD9⁺ EVs for the diagnosis of breast cancer (BC) but also in the quantitative analysis of CD14⁺ EVs to CD9⁺ EVs for the diagnosis of type 2 diabetes (T2DM). Collectively, our work provides new ideas for the diagnosis of breast cancer and type 2 diabetes while developing a versatile method for distinguishing other diseases by simultaneous detection of multibiomarkers of Exos subtypes in trace samples due to the substitutability of antibodies.

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来源期刊

Analytical Chemistry 化学-分析化学

CiteScore

12.10

自引率

12.20%

发文量

1949

审稿时长

1.4 months

期刊介绍： Analytical Chemistry, a peer-reviewed research journal, focuses on disseminating new and original knowledge across all branches of analytical chemistry. Fundamental articles may explore general principles of chemical measurement science and need not directly address existing or potential analytical methodology. They can be entirely theoretical or report experimental results. Contributions may cover various phases of analytical operations, including sampling, bioanalysis, electrochemistry, mass spectrometry, microscale and nanoscale systems, environmental analysis, separations, spectroscopy, chemical reactions and selectivity, instrumentation, imaging, surface analysis, and data processing. Papers discussing known analytical methods should present a significant, original application of the method, a notable improvement, or results on an important analyte.