The article reviews the advancements in single-cell long-read sequencing technologies, highlighting their applications and challenges. Long-read sequencing technologies, such as PacBio and Oxford Nanopore, have significantly improved the accuracy and resolution of transcriptome, genome, and epigenome sequencing at the single-cell level. These technologies enable the capture of full-length transcripts, facilitating the identification of complex splicing patterns and novel isoforms. The review discusses the integration of long-read sequencing into existing single-cell RNA sequencing (scRNA-seq) pipelines, emphasizing the importance of accurate assignment of cell barcodes (CBs) and unique molecular identifiers (UMIs). It also covers the choice of long-read sequencing platforms, data analysis pipelines, and specific applications in various fields, including developmental biology, cancer research, and spatial transcriptomics. The article identifies key challenges, such as the lack of streamlined library preparation protocols, the detection of rare isoforms, and the need for comparative benchmarking of bioinformatics tools. Finally, it outlines future directions for advancing single-cell long-read sequencing technologies to better understand cellular heterogeneity and function.The article reviews the advancements in single-cell long-read sequencing technologies, highlighting their applications and challenges. Long-read sequencing technologies, such as PacBio and Oxford Nanopore, have significantly improved the accuracy and resolution of transcriptome, genome, and epigenome sequencing at the single-cell level. These technologies enable the capture of full-length transcripts, facilitating the identification of complex splicing patterns and novel isoforms. The review discusses the integration of long-read sequencing into existing single-cell RNA sequencing (scRNA-seq) pipelines, emphasizing the importance of accurate assignment of cell barcodes (CBs) and unique molecular identifiers (UMIs). It also covers the choice of long-read sequencing platforms, data analysis pipelines, and specific applications in various fields, including developmental biology, cancer research, and spatial transcriptomics. The article identifies key challenges, such as the lack of streamlined library preparation protocols, the detection of rare isoforms, and the need for comparative benchmarking of bioinformatics tools. Finally, it outlines future directions for advancing single-cell long-read sequencing technologies to better understand cellular heterogeneity and function.