Tissue preparation for high-quality RNA retrieval via Laser Capture Microdissection (LCM)

Wednesday, October 26, 2011 — Poster Session III
10:00 a.m. – Noon	Natcher Conference Center	OD	TECH-4

Authors

• Y Golubeva
• R Smith
• L Sternberg

Abstract

Laser Capture Microdissection (LCM) is a preferred method for sample enrichment prior to RNA expression analysis. Successful LCM depends on optimal sample preparation, inactivation of endogenous nucleases and protection from exogenous compromise of target RNA. Accordingly, the majority of LCM projects are conducted on fresh frozen tissue that contain native (unmodified by fixation) nucleic acid. In the absence of fixation however, the physical characteristics of tissues that contain significant fat or collagen or with especially delicate anatomical structures create significant additional complications to successful LCM. To address this specialized challenge within LCM, we adapted the CryoJane Tape-Transfer System (Leica, USA), an existing solution for efficient transfer of frozen tissue sections to glass microscope slides for use with LCM (glass) and Laser Cutting (membrane) slides. Modification to the GryoJane coating protocol for the membrane slides facilitated tissue transfer on even the most challenging of frozen tissues. We next developed and validated companion protocols that facilitated efficient dissection and high quality RNA retrieval (RIN ≥ 7.0) from the modified CryoJane preparation.

back to top