Linked Life Data

20888418

Source:http://linkedlifedata.com/resource/pubmed/id/20888418

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
4
pubmed:dateCreated
2010-11-18
pubmed:abstractText
Clinical trials and correlative laboratory research are increasingly reliant upon archived paraffin-embedded samples. Therefore, the proper processing of biological samples is an important step to sample preservation and for downstream analyses like the detection of a wide variety of targets including micro RNA, DNA and proteins. This paper analyzed the question whether routine fixation of cells and tissues in 10% buffered formalin is optimal for in situ and solution phase analyses by comparing this fixative to a variety of cross linking and alcohol (denaturing) fixatives. We examined the ability of nine commonly used fixative regimens to preserve cell morphology and DNA/RNA/protein quality for these applications. Epstein-Barr virus (EBV) and bovine papillomavirus (BPV)-infected tissues and cells were used as our model systems. Our evaluation showed that the optimal fixative in cell preparations for molecular hybridization techniques was "gentle" fixative with a cross-linker such as paraformaldehyde or a short incubation in 10% buffered formalin. The optimal fixatives for tissue were either paraformaldehyde or low concentration of formalin (5% of formalin). Methanol was the best of the non cross-linking fixatives for in situ hybridization and immunohistochemistry. For PCR-based detection of DNA or RNA, some denaturing fixatives like acetone and methanol as well as "gentle" cross-linking fixatives like paraformaldehyde out-performed other fixatives. Long term fixation was not proposed for DNA/RNA-based assays. The typical long-term fixation of cells and tissues in 10% buffered formalin is not optimal for combined analyses by in situ hybridization, immunohistochemistry, or--if one does not have unfixed tissues--solution phase PCR. Rather, we recommend short term less intense cross linking fixation if one wishes to use the same cells/tissue for in situ hybridization, immunohistochemistry, and solution phase PCR.
pubmed:grant
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Dec
pubmed:issn
1095-9130
pubmed:author
pubmed:copyrightInfo
Published by Elsevier Inc.
pubmed:issnType
Electronic
pubmed:volume
52
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
281-6
pubmed:meshHeading
pubmed-meshheading:20888418-Animals, pubmed-meshheading:20888418-Cattle, pubmed-meshheading:20888418-Cell Line, pubmed-meshheading:20888418-DNA, Viral, pubmed-meshheading:20888418-Fixatives, pubmed-meshheading:20888418-Formaldehyde, pubmed-meshheading:20888418-Herpesvirus 4, Human, pubmed-meshheading:20888418-Humans, pubmed-meshheading:20888418-Immunohistochemistry, pubmed-meshheading:20888418-In Situ Hybridization, pubmed-meshheading:20888418-MicroRNAs, pubmed-meshheading:20888418-Nucleic Acid Hybridization, pubmed-meshheading:20888418-Papillomaviridae, pubmed-meshheading:20888418-Paraffin Embedding, pubmed-meshheading:20888418-Polymerase Chain Reaction, pubmed-meshheading:20888418-Polymers, pubmed-meshheading:20888418-Preservation, Biological, pubmed-meshheading:20888418-RNA, pubmed-meshheading:20888418-Tissue Fixation
pubmed:year
2010
pubmed:articleTitle
The search for an optimal DNA, RNA, and protein detection by in situ hybridization, immunohistochemistry, and solution-based methods.
pubmed:affiliation
Divisions of Hematology & Medical Oncology, Department of Internal Medicine, Comprehensive Cancer Center, College of Medicine, The Ohio State University, 320 West 10th Avenue, Columbus, OH 43210, USA.
pubmed:publicationType
Journal Article, Research Support, N.I.H., Extramural