Cell membrane nanostructure is altered by heat-induced antigen retrieval:A possible consequence for immunocytochemical detection of membranous antigens

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F61989592%3A15110%2F20%3A73599480" target="_blank" >RIV/61989592:15110/20:73599480 - isvavai.cz</a>

Výsledek na webu

<a href="https://www.cambridge.org/core/journals/microscopy-and-microanalysis/article/abs/cell-membrane-nanostructure-is-altered-by-heatinduced-antigen-retrieval-a-possible-consequence-for-immunocytochemical-detection-of-membranous-antigens/2B3F902B8BD819D22B91F8D08718A41D" target="_blank" >https://www.cambridge.org/core/journals/microscopy-and-microanalysis/article/abs/cell-membrane-nanostructure-is-altered-by-heatinduced-antigen-retrieval-a-possible-consequence-for-immunocytochemical-detection-of-membranous-antigens/2B3F902B8BD819D22B91F8D08718A41D</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1017/S1431927619015113" target="_blank" >10.1017/S1431927619015113</a>

Jazyk výsledku

angličtina

Název v původním jazyce

Cell membrane nanostructure is altered by heat-induced antigen retrieval:A possible consequence for immunocytochemical detection of membranous antigens

Popis výsledku v původním jazyce

Heat-induced antigen retrieval (HIAR) improves immunodetection of antigens in formalin-fixed, paraffin-embedded tissue samples and it can also improve detection especially of intracellular antigens in alcohol-fixed cytological samples although it can have deleterious effect especially for membranous antigens. We examined the differences in cell surface topography by atomic force microscopy on methanol:aceton- (M:A) and 4% paraformaldehyde (4% PFA)-fixed MCF7 cells and changes caused by HIAR at three different temperatures (60, 90, 120°C). Moreover, the consequences for immunostaining of five membranous antigens (EGFR, E-cadherin, CD9, CD24 and CD44) were examinated. Our result showed that albeit there is no optimal immunostaining condition for our tested antibodies, the surface topography could be one of the important factors. Generally, the best conditions for successful immunostaining are M:A fixation with no HIAR treatment whereas in 4% PFA-fixed cells HIAR 120 is optimal. These conditions showed similarity in cell surface skewness which indicates with correlation factor 0.8000 between successful ICC staining and skewness parameter. It indicates that presence of valleys, depressions, scratches and pits on cell surface represents unfavourable condition for successful immunodetection of cell surface antigens.

Název v anglickém jazyce

Cell membrane nanostructure is altered by heat-induced antigen retrieval:A possible consequence for immunocytochemical detection of membranous antigens

Popis výsledku anglicky

Heat-induced antigen retrieval (HIAR) improves immunodetection of antigens in formalin-fixed, paraffin-embedded tissue samples and it can also improve detection especially of intracellular antigens in alcohol-fixed cytological samples although it can have deleterious effect especially for membranous antigens. We examined the differences in cell surface topography by atomic force microscopy on methanol:aceton- (M:A) and 4% paraformaldehyde (4% PFA)-fixed MCF7 cells and changes caused by HIAR at three different temperatures (60, 90, 120°C). Moreover, the consequences for immunostaining of five membranous antigens (EGFR, E-cadherin, CD9, CD24 and CD44) were examinated. Our result showed that albeit there is no optimal immunostaining condition for our tested antibodies, the surface topography could be one of the important factors. Generally, the best conditions for successful immunostaining are M:A fixation with no HIAR treatment whereas in 4% PFA-fixed cells HIAR 120 is optimal. These conditions showed similarity in cell surface skewness which indicates with correlation factor 0.8000 between successful ICC staining and skewness parameter. It indicates that presence of valleys, depressions, scratches and pits on cell surface represents unfavourable condition for successful immunodetection of cell surface antigens.

Druh

J_imp - Článek v periodiku v databázi Web of Science

CEP obor

—

OECD FORD obor

30106 - Anatomy and morphology (plant science to be 1.6)

Projekt

Výsledek vznikl pri realizaci vícero projektů. Více informací v záložce Projekty.

Návaznosti

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Rok uplatnění

2020

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název periodika

Microscopy and Microanalysis

ISSN

1431-9276

e-ISSN

—

Svazek periodika

26

Číslo periodika v rámci svazku

1

Stát vydavatele periodika

US - Spojené státy americké

Počet stran výsledku

9

Strana od-do

139-147

Kód UT WoS článku

000516841400017

EID výsledku v databázi Scopus

2-s2.0-85080827035