基础医学与临床2026，Vol.46Issue(7)：895-903,9.DOI:10.16352/j.issn.1001-6325.2026.07.0895
理论光谱在小鼠骨髓细胞光谱流式细胞测量术中的应用
Application of theoretical spectra in spectral flow cytometry of mouse bone marrow cells
摘要
Abstract
Objective To investigate the feasibility and limitations of theoretical spectra applied as substitutes for single-stained controls in spectral flow cytometry,and to propose a reference-spectrum optimization strategy for correcting deviations between theoretical and experimentally measured spectra caused by auto-fluorescence.Methods Mouse bone marrow single-stained samples were collected and analyzed by multi-platform spectral flow cytometry.Cosine similarity was used to evaluate the consistency between theoretical spectra and experimentally measured spectra.For single-stained samples affected by autofluorescence,reference spectra were generated from cell subsets with relatively homogeneous auto-fluorescence,and their effects on performance were assessed.Results Among eleven antibody-fluorochromes/fluorescent dyes,five of them showed cosine similarity values of 0.90 or higher between measured and theoretical spectra on both platforms,which suggested that they might serve as alternatives to single-stained controls.Spectral mismatches manifested either as spectral shifts related to detec-tion conditions or,in a subset of samples,as abnormal reference spectra associated with auto-fluorescence inter-ference.The subset strategy corrected abnormal reference spectra caused by auto-fluorescence heterogeneity and significantly reduced unmixing spillover errors in mouse bone marrow cells.Conclusions Theoretical spectra can serve as effective tools for evaluating and correcting reference spectra and may partially replace single-stained con-trols.The subset strategy can reduce reference-spectrum bias caused by autofluorescence and improve the accuracy of spectral unmixing.关键词
光谱流式细胞测量术/理论光谱/参考光谱/自发荧光/预设子集策略Key words
spectral flow cytometry/theoretical spectra/reference spectra/autofluorescence/subset strategy分类
医药卫生引用本文复制引用
李卓,刘雪会,李静,范宏,樊化林,吕湘..理论光谱在小鼠骨髓细胞光谱流式细胞测量术中的应用[J].基础医学与临床,2026,46(7):895-903,9.基金项目
国家自然科学基金(82570165) （82570165）
疑难重症及罕见病全国重点实验室自主课题(2025-I-PY-004) （2025-I-PY-004）
中国医学科学院创新工程攻关项目(2025-I2M-KJ-017) （2025-I2M-KJ-017）
