Work

Enhanced RNA Preservation in Mouse Brain Tissue: A Strategy Combining Cardiac Perfusion with Hypersaline Immersion

Year: 2025

Type: article

Abstract: High Resolution Image Download MS PowerPoint Slide One of the key research methods for low-density spatial transcriptomics involves collecting and sequencing microregions of interest from tissue sections. However, prolonged capillary microneedle sampling poses a risk of RNA degradation. To address this issue, we developed a preservation strategy consisting of: (1) mouse cardiac perfusion with phosphate-buffered saline (PBS) to reduce blood-borne RNase activity; (2) brain tissue immersion in a hypersaline solution prior to sectioning; and (3) microregion sampling under low-temperature conditions. Assessment of tissue section RNA integrity and derived microregion transcriptomes demonstrated that this combined approach effectively preserves RNA integrity, maintaining an RNA integrity number equivalent greater than 7.5 for 6 h at 25 °C and greater than 8.5 for 24 h at 4 °C. Furthermore, this protective method reduced 3′ transcript bias and enhanced the detection of cell-type marker genes compared to controls ( p < 0.05). This strategy establishes a standardized framework to minimize preanalytical variability in capillary-based spatial omics studies of mouse brain sections.

Source: ACS Omega

DOI: 10.1021/acsomega.5c05379

Authors: Qinyu Ge, Erteng Jia, Tian Wen, Ruibin Zhao, Xiangwei Zhao

Institutions: State Key Laboratory of Digital Medical Engineering, Southeast University, Xuzhou Medical College, Jiangsu Provincial Center for Disease Control and Prevention

Language: English

Topic: Single-cell and spatial transcriptomics

Subfield: Molecular Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

SDG: Good health and well-being

Open Access status: gold

Estimated APC paid: $1,935