Non-small cell lung cancer(NSCLC) is one of the most common malignancies worldwide. Not only the complex molecular components but also the cellular heterogeneity in NSCLC tissues pose a great barrier for its clinical treatment. Recent years has witnessed the widespread application of single-cell sequencing in the studies regarding tumor heterogeneity. However, the disadvantages of single-cell sequencing technology itself could not be neglected. Spatial transcriptome(ST) technology allows in situ transcriptome sequencing of tissues to achieve high-throughput transcriptomic information of tissue cells with their spatial information available. In other words, ST makes it possible to acquire cellular composition and gene expression patterns without breaking intercellular communication network, which distinguishes itself from conventional single-cell sequencing, since mechanical separation and enzymatic digestion of tissue cells into single-cell suspension used to be inevitable during the performance of single-cell sequencing. To gain new insights into the spatial heterogeneity of NSCLC, we reviewed and summarized the latest progress in ST technology which has been applied to tumor sample analysis, especially to the field of NSCLC.