A nondestructive molecule extraction method allowing morphological and molecular analyses using a single tissue section.
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.
Chu
Wei-Sing
WS
Department of Scientific Laboratories, Armed Forces Institute of Pathology, Washington, DC 20306-6000, USA. chu@afip.osd.mil
Liang
Qi
Q
Liu
Jilan
J
Wei
Min Qi
MQ
Winters
Mary
M
Liotta
Lance
L
Sandberg
Glenn
G
Gong
Maokai
M
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
D002021:Buffers; D009696:Nucleic Acids; D011506:Proteins