This study explores the feasibility of single-cell proteomics in plant samples, focusing on *Arabidopsis thaliana* roots. The researchers isolated single cells from the cortex and endodermis, two adjacent root cell types derived from a common stem cell. From 756 cells, they identified 3,751 proteins and quantified 1,114 proteins per cell. After stringent filtering, 3,101 proteins were quantified, and 555 proteins were found to be enriched in either the cortex or endodermis, allowing for the differentiation of these closely related cell types. The findings highlight the potential of single-cell proteomics to explore the heterogeneity of expression between individual plant cells, providing insights into the spatiotemporal response to various biological stimuli.This study explores the feasibility of single-cell proteomics in plant samples, focusing on *Arabidopsis thaliana* roots. The researchers isolated single cells from the cortex and endodermis, two adjacent root cell types derived from a common stem cell. From 756 cells, they identified 3,751 proteins and quantified 1,114 proteins per cell. After stringent filtering, 3,101 proteins were quantified, and 555 proteins were found to be enriched in either the cortex or endodermis, allowing for the differentiation of these closely related cell types. The findings highlight the potential of single-cell proteomics to explore the heterogeneity of expression between individual plant cells, providing insights into the spatiotemporal response to various biological stimuli.