A Review of Technical Advances and Applications of High-DimensionalFlow Cytometry in Hematology
ZHOU Yidan, LIANG Xingguang*
Flow cytometry is a technique for multi-parameter detection, rapid quantitative analysis andphysical sorting of single cells or biological particles in cell populations. Accurate characterization of cellular immunophenotypes, which largely depends on the technical evolution and innovation of flow cytometry, is the coreprerequisite for deciphering the pathogenesis of hematological diseases. Conventional flow cytometry has inherentdrawbacks, including a limited number of detectable fluorescence channels, severe spectral overlap and low analytical dimensionality, rendering it insufficient for the analysis of highly complex biological specimens. Against thisbackground, three high-dimensional flow cytometry platforms, namely spectral flow cytometry, mass cytometryand imaging flow cytometry, have emerged, bringing revolutionary advances to the field of flow cytometry. Spectral flow cytometry deconvolves the full emission spectra of fluorescent dyes through dedicated algorithms, whicheffectively enhances the resolution of overlapping fluorescent signals and enables high-precision detection of over50 fluorescence parameters. Mass cytometry replaces conventional fluorescent dyes with heavy metal isotopeconjugated antibodies, completely avoiding spectral overlap artifacts. Imaging flow cytometry combines highthroughput cell profiling with microscopic imaging. It simultaneously acquires cellular fluorescence intensity, lightscatter signals and morphological features to achieve visualized multi-dimensional single-cell analysis. This reviewsystematically elaborates the basic principles and core performance indicators of the above three high-dimensionalflow cytometry modalities, focusing on their state-of-the-art applications in hematological disorders. It aims to offerobjective technical references for researchers to select appropriate high-dimensional flow cytometry platforms fordifferent experimental and clinical contexts.
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