Researchers from the RIKEN Cluster for Pioneering Research have developed a microfluidic method for electrical lysis and RNA extraction from single fixed cells leveraging reversible cross-linker dithiobis(succinimidyl propionate) (DSP). The microfluidic system captures a single DSP-fixed cell at a hydrodynamic trap, reverse-cross-links the DSP molecules on a chip with dithiothreitol, lyses the plasma membrane via electrical field, and extracts cytoplasmic RNA with isotachophoresis-aided nucleic acids extraction. All of the on-chip processes complete in less than 5 min. The researchers demonstrated the method using K562 leukemia cells and benchmarked the performance of RNA extraction with reverse transcription quantitative polymerase chain reaction. They also demonstrated the integration of our method with single-cell RNA sequencing.
Electrical lysis and RNA extraction from single cells for RNA sequencing
Subramanian Parimalam S, Oguchi Y, Abdelmoez MN, Tsuchida A, Ozaki Y, Yokokawa R, Kotera H, Shintaku H. (2018) Electrical Lysis and RNA Extraction from Single Cells Fixed by Dithiobis(succinimidyl propionate). Anal Chem [Epub ahead of print]. [abstract]