title: A nondestructive molecule extraction method allowing morphological and molecular analyses using a single tissue section.
abstract: In clinical practice, molecular analysis of tumor specimens is often restricted by available technology for sample preparation. Virtually all current methods require homogenization of tissues for molecule extraction. We have developed a simple, rapid, nondestructive molecule extraction (NDME) method to extract proteins and nucleic acids directly from a single fixed or frozen tissue section without destroying the tissue morphology. The NDME method is based upon exposure of micron-thick tissue section to extraction buffer with the help of heating and/or intact physical forces (ultrasound and microwave) to facilitate release of macromolecules into the buffer. The extracted proteins and nucleic acids can be used directly without further purification for downstream SDS-PAGE analysis, immunoblotting, protein array, mass spectra protein profiling, PCR, and RT-PCR reactions. Most importantly, the NDME procedure also serves as an antigen retrieval treatment, so that after NDME, the same tissue section can be used for histopathological analyses, such as H&E staining, immunohistochemistry, and in situ hybridization. Thus, the NDME method allows, for the first time, both histological diagnosis and molecular analysis on a single tissue section, whether it is from frozen or fixed tissue specimens.
authors.affiliation: Department of Scientific Laboratories, Armed Forces Institute of Pathology, Washington, DC 20306-6000, USA. chu@afip.osd.mil
authors.forename: Wei-Sing
authors.initials: WS
authors.lastname: Chu
authors.forename: Qi
authors.initials: Q
authors.lastname: Liang
authors.forename: Jilan
authors.initials: J
authors.lastname: Liu
authors.forename: Min Qi
authors.initials: MQ
authors.lastname: Wei
authors.forename: Mary
authors.initials: M
authors.lastname: Winters
authors.forename: Lance
authors.initials: L
authors.lastname: Liotta
authors.forename: Glenn
authors.initials: G
authors.lastname: Sandberg
authors.forename: Maokai
authors.initials: M
authors.lastname: Gong
mesh_terms: D000818:Animals; D001709:Biotechnology; D002021:Buffers; D005629:Frozen Sections; D006358:Hot Temperature; D006801:Humans; D007150:Immunohistochemistry; D008867:Microtomy; D008872:Microwaves; D009696:Nucleic Acids / Q000032:analysis* / Q000302:isolation & purification*; D016612:Paraffin Embedding; D011506:Proteins / Q000032:analysis* / Q000302:isolation & purification*; D016707:Tissue Fixation; D014465:Ultrasonics
chemical_list: D002021:Buffers; D009696:Nucleic Acids; D011506:Proteins