Beneficial effect of heat-induced antigen retrieval in immunocytochemical detection of intracellular antigens in alcohol-fixed cell samples
The result's identifiers
Result code in IS VaVaI
<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F61989592%3A15110%2F20%3A73591176" target="_blank" >RIV/61989592:15110/20:73591176 - isvavai.cz</a>
Result on the web
<a href="https://journals.lww.com/appliedimmunohist/Abstract/2020/02000/Beneficial_Effect_of_Heat_induced_Antigen.11.aspx" target="_blank" >https://journals.lww.com/appliedimmunohist/Abstract/2020/02000/Beneficial_Effect_of_Heat_induced_Antigen.11.aspx</a>
DOI - Digital Object Identifier
<a href="http://dx.doi.org/10.1097/PAI.0000000000000689" target="_blank" >10.1097/PAI.0000000000000689</a>
Alternative languages
Result language
angličtina
Original language name
Beneficial effect of heat-induced antigen retrieval in immunocytochemical detection of intracellular antigens in alcohol-fixed cell samples
Original language description
Immunohistochemistry (IHC) and immunocytochemistry (ICC) play an irreplaceable role in research and diagnostics. It is well known that antigen retrieval (AR) can, as a technique, have beneficial outcomes on IHC results when using formalin-fixed, paraffin-embedded tissue samples. The main purpose of AR is to break protein crosslinks which are formed during formalin fixation. Although AR was originally designed for FFPE samples, the usefulness of AR in ICC has been described. Cytological samples are often fixed in alcohol-based fixatives which does not lead to the formation of crosslinks. Therefore, alcohol-fixed samples can be successfully immunostained without AR. We investigated the effect of heat-induced antigen retrieval (HIAR) on alcohol-fixed HEK293 cell line samples and patient cytological samples from thyroid gland obtained by fine needle aspiration (FNA) technique. We compared indirect two-step ICC staining results performed according to the protocol with or without HIAR in citrate buffer pH6 for several antibodies. Utilising HIAR against intracellular antigens has beneficial effects. Therefore, more diluted antibodies can be used for satisfactory results. However, surface antigens were probably damaged by HIAR treatment. We demonstrated evident changes in cell surface topography after HIAR treatment by atomic force microscopy. Staining specificity of patient samples improves and background staining is reduced, allowing higher dilutions of primary antibody. Improving staining specificity is necessary for accurate diagnostics. Although we have shown the beneficial effect of HIAR for immunostaining intracellular antigens, proper staining protocol should be tested on appropriate controls for individual antibodies.
Czech name
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Czech description
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Classification
Type
J<sub>imp</sub> - Article in a specialist periodical, which is included in the Web of Science database
CEP classification
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OECD FORD branch
30106 - Anatomy and morphology (plant science to be 1.6)
Result continuities
Project
Result was created during the realization of more than one project. More information in the Projects tab.
Continuities
P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)
Others
Publication year
2020
Confidentiality
S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů
Data specific for result type
Name of the periodical
APPLIED IMMUNOHISTOCHEMISTRY & MOLECULAR MORPHOLOGY
ISSN
1541-2016
e-ISSN
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Volume of the periodical
28
Issue of the periodical within the volume
2
Country of publishing house
US - UNITED STATES
Number of pages
9
Pages from-to
166-174
UT code for WoS article
000516601200014
EID of the result in the Scopus database
2-s2.0-85052788339