DRaqL Method Allows Precise Single-Cell Transcriptome Analysis from Tissue Sections

A novel method called DRaqL (direct RNA recovery and quenching for laser capture microdissection) has been developed by researchers from Nara Medical University. According to a paper published in Life Science Alliance, DRaqL enables highly sensitive quantitative single-cell transcriptome analysis, providing precise insights into cells extracted from tissue sections using laser capture microdissection (LCM).

This innovative technique also facilitates quantification of exon-exon junctions of mRNA. When applied to mouse ovarian sections, DRaqL allowed the creation of a predictive model for oocyte transcriptomes based on their diameter. Deviations from this model unveiled growth retardation in oocyte gene expression profiles. Additionally, the research revealed gene expression differences in granulosa cells depending on their proximity to oocytes.

This study marks the first comprehensive quantitative analysis of the relationship between transcriptomes and histology during folliculogenesis at the single-cell level. It was conducted in collaboration with Kyoto University and RIKEN.

DRaqL overcomes the limitations of traditional methods by efficiently lysing cells isolated from alcohol- or formalin-fixed tissue sections and applying the lysate directly to RNA sequencing. This method demonstrates sensitivity comparable to single-cell RNA-seq from freshly isolated cells, even when used on strongly fixed, formalin-treated tissue sections.

The capacity of DRaqL to enable high-quality single-cell transcriptome analysis opens up new possibilities for understanding the relationship between morphology and gene expression in various biological contexts. This method has immense potential in fertility treatment, assisted reproductive medicine, and other fields, offering insights into the quality control of follicles and oocytes.