DroNc-seq is a massively parallel single nucleus RNA-seq method that enables high-throughput, sensitive, and unbiased classification of cell types from archived brain tissues. The method combines the advantages of sNuc-Seq and Drop-seq, allowing for efficient profiling of nuclei with low cost and high throughput. The study demonstrates the application of DroNc-seq on mouse and human archived brain samples, revealing distinct cell types and subtypes, rare cells, expression signatures, and activated pathways. The method successfully identifies cell-type specific gene signatures, which are consistent with previously defined signatures in mouse and human tissues. DroNc-seq is robust, cost-effective, and easy to use, enabling the systematic analysis of complex tissues that are difficult to dissociate or already archived. The method is particularly useful for analyzing archived human tissues, such as post-mortem brain samples, and can identify rare cell types without the need for enrichment. The study also shows that DroNc-seq provides comparable results to other single-cell and single-nucleus RNA-seq methods, with improved throughput and cost. The method is applicable to a wide range of tissues and can be used to create vital atlases of human tissues and clinical samples. The study highlights the potential of DroNc-seq for advancing our understanding of cell types and their functions in complex tissues.DroNc-seq is a massively parallel single nucleus RNA-seq method that enables high-throughput, sensitive, and unbiased classification of cell types from archived brain tissues. The method combines the advantages of sNuc-Seq and Drop-seq, allowing for efficient profiling of nuclei with low cost and high throughput. The study demonstrates the application of DroNc-seq on mouse and human archived brain samples, revealing distinct cell types and subtypes, rare cells, expression signatures, and activated pathways. The method successfully identifies cell-type specific gene signatures, which are consistent with previously defined signatures in mouse and human tissues. DroNc-seq is robust, cost-effective, and easy to use, enabling the systematic analysis of complex tissues that are difficult to dissociate or already archived. The method is particularly useful for analyzing archived human tissues, such as post-mortem brain samples, and can identify rare cell types without the need for enrichment. The study also shows that DroNc-seq provides comparable results to other single-cell and single-nucleus RNA-seq methods, with improved throughput and cost. The method is applicable to a wide range of tissues and can be used to create vital atlases of human tissues and clinical samples. The study highlights the potential of DroNc-seq for advancing our understanding of cell types and their functions in complex tissues.