RNA sampling from tissue sections using infrared laser ablation
RNA was obtained from discrete locations of frozen rat brain tissue sections through infrared (IR) laser ablation using a 3-μm wavelength in transmission geometry. The ablated plume was captured in a microcentrifuge tube containing RNAse-free buffer and processed using a commercial RNA purification kit. RNA transfer efficiency and integrity were evaluated based on automated electrophoresis in microfluidic chips. Reproducible IR-laser ablation of intact RNA was demonstrated with purified RNA at laser fluences of 3-5 kJ/m (72 ± 12% transfer efficiency) and with tissue sections at a laser fluence of 13 kJ/m (79 ± 14% transfer efficiency); laser energies were attenuated ∼20% by the soda-lime glass slides used to support the samples. RNA integrity from tissue ablation was >90% of its original RIN value (∼7) and the purified RNA was sufficiently intact for conversion to cDNA and subsequent qPCR assay.
Publication Source (Journal or Book title)
Analytica chimica acta
Wang, K., Donnarumma, F., Herke, S. W., Dong, C., Herke, P. F., & Murray, K. K. (2019). RNA sampling from tissue sections using infrared laser ablation. Analytica chimica acta, 1063, 91-98. https://doi.org/10.1016/j.aca.2019.02.054