In situ hybridization in human and rodent tissue by the use of a new and simplified method.
In situ hybridization (ISH) is a method that detects and localizes DNA or RNA in morphologically preserved tissue and cell preparations. The method is based on the principle that DNA or RNA will undergo hydrogen binding to complimentary sequences. Selective probes are labeled and used in order to detect specific sequences in tissues or cell preparations. Even though the method has improved over the past decades, there are still issues with sensitivity and specificity. The protocols are nonstandardized, and often time consuming due to multiple steps. In this paper, we have used a new and commercially available ISH kit for the detection of mRNA in formalin-fixed paraffin-embedded tissue. We have used both human and Mongolian gerbil tissue, and we evaluated mRNA expression of the neuroendocrine markers chromogranin A and histidine decarboxylase in both normal tissue and poorly differentiated tumor. In our experience, this method offers excellent sensitivity and specificity. The protocol is more standardized, and our results have been consistent. It is also less time consuming than conventional ISH protocols.
Department of Cancer Research and Molecular Medicine, Norwegian University of Science and Technology (NTNU), Trondheim, Norway. oystein.sordal@ntnu.no
Øystein
Ø
Sørdal
Gunnar
G
Qvigstad
Ivar Skjåk
IS
Nordrum
Bjørn
B
Gustafsson
Helge L
HL
Waldum
D000818:Animals; D002277:Carcinoma / Q000175:diagnosis* / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D053379:Chromogranin A / Q000235:genetics* / Q000378:metabolism; D005557:Formaldehyde; D015870:Gene Expression; D005849:Gerbillinae; D016481:Helicobacter Infections / Q000175:diagnosis / Q000235:genetics / Q000382:microbiology / Q000473:pathology*; D006640:Histidine Decarboxylase / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D006965:Hyperplasia; D007150:Immunohistochemistry; D017403:In Situ Hybridization / Q000379:methods / Q000592:standards*; D016612:Paraffin Embedding; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics* / Q000378:metabolism; D011933:Reagent Kits, Diagnostic; D012680:Sensitivity and Specificity; D013274:Stomach Neoplasms / Q000175:diagnosis* / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D016707:Tissue Fixation
D053379:Chromogranin A; D012333:RNA, Messenger; D011933:Reagent Kits, Diagnostic; D005557:Formaldehyde; D006640:Histidine Decarboxylase