Cytopathological evaluations combined RNA and protein analyses on defined cell regions using single frozen tissue block

Laboratory of Cell Biology and Molecular Genetics, College of Basic Medical Sciences, Dalian Medical University, Dalian 116027, China

The co-existence of multiple cell components in tissue samples is the main obstacle for precise molecular evaluation on defined cell types. Based on morphological examination, we developed an efficient approach for paralleled RNA and protein isolations from an identical histological region in frozen tissue section. The RNA and protein samples prepared were sufficient for RT-PCR and Western blot analyses, and the results obtained were well coincident each other as well as with the corresponding parameters revealed from immunohistochemical examinations. By this way, the sampling problem caused by cell-cross contamination can be largely avoided, committing the experimental data more specific to a defined cell type. These novel methods thus allow us to use single tissue block for a comprehensive study by integration of conventional cytological evaluations with nucleic acid and protein analyses.

Key words: RNA and protein preparation, defined histological region, serial frozen section, gene expression.