“…Single-cell RNA-sequencing (scRNA-seq) technology has been widely used in many biological investigations, including the elucidation of cell subtype heterogeneity ( Zeisel et al 2015 ; Goolam et al 2016 ), construction of gene regulatory networks ( Darmanis et al 2015 ), profiling of cell development and differentiation ( Deng et al 2014 ; Liu et al 2017 ), and depiction of disease in an immunoresponsive environment ( Guo et al 2018 ; Zhang et al 2018 ). The analysis of scRNA-seq data contains, but is not limited to, quality control ( Chen et al 2016 ), data normalization ( Cole et al 2019 ), unsupervised clustering ( Kiselev et al 2017 ; Wang et al 2017 ; Wolf et al 2018 ; Yang and Wang 2020 ), trajectory construction ( Wolf et al 2019 ), and differentially expressed gene identification ( Soneson and Robinson 2018 ). As a fundamental step of scRNA-seq data analysis, cell clustering determines the results of subsequent downstream analyses to a certain extent, but is often inaccurate and misconstrues analyses.…”