RNA　splicing　is　a　fundamental　regulatory　process　of　gene　expression　and　plays　a　pivotal　role　in　transcriptome　complexity,　cell-fate　determination,　and　organism　development.　The　detection　of　splicing　products　has　attracted　significant　interest　because　aberrant　splicing　can　lead　to　cell　dysfunction　and　numerous　diseases,　such　as　cancer　and　neurodegeneration.　Particularly,　splicing　variability　between　individual　cells　is　primarily　responsible　for　gene　expression　heterogeneity.　Investigations　into　single-cell　expression　of　RNA　splicing　variants　offer　a　venue　for　resolving　gene　regulatory　circuits,　mapping　intracellular　localization,　and　classifying　cell　types.　This　review　discusses　the　merits　and　technical　challenges　in　transitioning　RNA　splicing　detection　from　in　vitro　testing　to　single-cell　analysis.　In　the　end,　we　suggest　some　future　directions　for　RNA　splicing　analysis　and　expect　to　inspire　innovative　works　and　discoveries　in　this　field.