Zobrazovací imunohistochemická metoda pro značení řezů sferoidů s využitím zlatých nanočástic pomocí laserové ablace a hmotnostní spektrometrie s indukčně vázaným plazmatem
| Title in English | Imaging immunohistochemical method for labeling spheroid sections using gold nanoparticles by laser ablation and inductively coupled plasma mass spectrometry |
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| Authors | |
| Year of publication | 2021 |
| Type | Conference abstract |
| MU Faculty or unit | |
| Citation | |
| Description | Spheroids are used 3D cell models that can represent the actual environment of the tumor tissue. Thus, they are a good model for immunohistochemical (IHC) labeling and 2D or 3D imaging of the target protein in tissue sections. The distribution of proliferating cells in the outer parts, where nutrients and oxygen are abundant, and the occurrence of necrotic cells closer to the center of the spheroid thus resemble the tumor microenvironment. One marker of proliferation is the Ki-67 protein, for the detection of which fluorescence microscopy is routinely used to label primary and secondary antibodies with a fluorophore. The formation of a very tight avidin-biotin bond is also often used, where one of the partners is labeled with a detection tag. Labelling the Ki-67 protein with a gold nanoparticle (AuNP) of defined size allows highly sensitive determination of this marker by laser ablation coupled to inductively coupled plasma mass spectrometry (LA ICP MS). In this paper, the development of an IHC labeling method for spheroid sections formed by colon adenocarcinoma cells from the HT-29 line is described. In particular, attention is paid to the elimination of non-specific nanoparticle interactions using protein or non-protein blocking solutions. 20nm AuNPs with adsorbed or covalently bound streptavidin are used. |
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