基于量子点编码微球的流式检测技术用于白血病诊断的研究

doi:10.3969/j.issn.1000-5641.2022.06.014

摘要/Abstract

摘要：

采用种子聚合法制备了微米级的聚苯乙烯微球 (Polystyrene Microspheres, PS), 并将微球羧基化和多孔化. 将量子点 (Quantum Dots, QDs) 作为荧光分子, 合成了不同发射波长的量子点, 并成功将其装载到多孔微球当中形成荧光编码微球 (QDs@PS), 最后检测急性白血病 (Acute Leukemia, AL) 患者血清样本, 通过流式实现对血清中抗原的定量分析. 扫描电子显微镜 (Scanning Electron Microscope, SEM) 和流式细胞术图像显示, 微球的球形规整, 尺寸均一. 荧光显微镜观察, QDs基本均匀渗透到整个微球中. 此外, QDs@PS显示了良好的光稳定性, 未见QDs泄漏, 并可保持其荧光至少两周. 荧光光谱分析对人免疫球蛋白G (Immunoglobulin G, IgG) 检测的性能表明, 荧光微球表面的羧基有利于生物大分子的高效共价结合, 可应用于三明治免疫夹心反应偶联白血病高表达抗原白介素6 (Interleukin 6, IL-6). 结合白血病患者血清样品, 通过流式细胞仪检测出量子点的荧光, 计算其平均荧光强度 (Mean Fluorescence Intensity, MFI) 来表示血清中IL-6的含量. 这些结果表明, 设计的光学编码微载体可以成功地应用于高通量和多路生物分子分析, 在血液疾病检测和诊断中有很大潜力.

关键词: 多孔聚苯乙烯微球, 量子点, 流式细胞术, 白血病, IL-6

Abstract:

In this work, micro-scale porous carboxyl polystyrene microspheres (PS) were prepared using seed polymerization. Quantum dots (QDs) were used as fluorescent molecules to synthesize QDs with different emission wavelengths and successfully loaded into porous microspheres to form fluorescent coding microspheres (QDs@PS). Subsequently, serum samples of patients with acute leukemia (AL) were detected, and the antigen in the serum was quantitatively analyzed using flow cytometry. Scanning electron microscope (SEM) and flow cytometry images showed that the microspheres were regular and uniform in size. Fluorescence microscopy showed that the QDs permeated uniformly into the whole microsphere. In addition, QDs@PS showed good fluorescence stability, no QD leakage was observed, and the QDs@PS maintained its fluorescence for a period of at least two weeks. The use of fluorescence spectroscopic analysis for the detection of human immunoglobulin G (IgG) showed that the carboxyl groups on the surface of fluorescent microspheres are beneficial for the efficient covalent binding of biological macromolecules, which can be used for sandwich immunosandwich reaction coupling with leukemic high expression antigen interleukin 6 (IL-6). Combined with serum samples from leukemia patients, the fluorescence of QDs was detected by flow cytometry, and the mean fluorescence intensity (MFI) was calculated to determine the content of IL-6 in the serum. These results indicate that the designed optically-encoded microcarrier can be successfully applied to high-throughput and multichannel biomolecular analysis and has great potential in blood disease detection and diagnosis.

Key words: carboxyl polystyrene microsphere, quantum dots, flow cytometry, leukemia, interleukin 6

中图分类号:

R730.43

杨俊飞, 王烨颖, 周靖娥, 王镜, 闫志强, 俞磊. 基于量子点编码微球的流式检测技术用于白血病诊断的研究[J]. 华东师范大学学报（自然科学版）, 2022, 2022(6): 139-149.

Junfei YANG, Yeying WANG, Jing’e ZHOU, Jing WANG, Zhiqiang YAN, Lei YU. Research on diagnosis of leukemia by flow detection based on quantum dot coded microspheres[J]. Journal of East China Normal University(Natural Science), 2022, 2022(6): 139-149.

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