中国激光, 2020, 47 (11): 1107001, 网络出版: 2020-11-02
基于显微镜数字图像的荧光免疫层析定量检测方法 下载: 853次
Quantitative Detection Method of Fluorescence Immunochromatography Based on Microscopic Digital Image
生物光学 光通量损失 数字图像处理 面阵CCD 定量检测 荧光免疫层析 biotechnology luminous flux loss digital image processing area array CCD quantitative detection fluorescence immunochromatography
摘要
荧光免疫层析定量检测技术已被广泛应用于临床检测领域,如新型冠状病毒肺炎的筛查和检测。为了提高检测精准度,提出了基于荧光显微镜数字图像处理的荧光免疫层析定量检测方法。首先构建了显微系统的光通量损失方程,然后根据面阵CCD输出的数字图像特征提取原始图像中的RGB分量并对其进行加权处理,之后拟合了面阵CCD的量子响应速率曲线,获得了荧光信号的相对强度。根据荧光信号强度即可反演出待测物的浓度。实验验证后可知,检测结果的变异系数为3.04%,线性拟合系数大于0.99。本检测方法可对最低质量浓度为0.1 ng/mL的待测物进行准确检测,适用于CCD成像型荧光显微镜系统,对荧光检测具有一定的参考意义。
Abstract
Fluorescence immunochromatographic quantitative detection technology has been widely used in the field of clinical detection, such as for the screening and detection of the new coronavirus pneumonia. To improve the detection accuracy, a fluorescence immunochromatographic quantitative detection method based on fluorescence microscope digital image processing is proposed. Initially, the luminous flux loss equation of the microscopy system is constructed. Then, the RGB components in the original image are extracted according to the digital image features output by the area array CCD, and the quantum response rate curve of the area array CCD is fitted after weighting to compensate. Finally, the relative intensity of the fluorescence signal is obtained, and the concentration of the analyte is inversed according to the intensity of the fluorescence signal. After experimental verification, it can be observed that the coefficient of variation of the test results is 3.04%, and the linear fitting coefficient is >0.99. This detection method can accurately detect the analyte with a minimum mass concentration of 0.1 ng/mL. It is suitable for CCD imaging fluorescent microscope system and has certain reference significance for fluorescence detection.
李婷婷, 冯大伟, 李琦, 向阳, 贾强, 路雨桐, 赵阳. 基于显微镜数字图像的荧光免疫层析定量检测方法[J]. 中国激光, 2020, 47(11): 1107001. Li Tingting, Feng Dawei, Li Qi, Xiang Yang, Jia Qiang, Lu Yutong, Zhao Yang. Quantitative Detection Method of Fluorescence Immunochromatography Based on Microscopic Digital Image[J]. Chinese Journal of Lasers, 2020, 47(11): 1107001.