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Bioconjugated and Cross-Linked Bionanostructures for Bifunctional Immunohistochemical Labeling

Published online by Cambridge University Press:  13 March 2012

Rıdvan Say*
Affiliation:
Department of Chemistry, Anadolu University, Eskişehir, Turkey
Gözde Aydoğan Kılıç
Affiliation:
Department of Biology, Anadolu University, Eskişehir, Turkey
Ayça Atılır Özcan
Affiliation:
Department of Chemistry, Anadolu University, Eskişehir, Turkey
Deniz Hür
Affiliation:
Department of Chemistry, Anadolu University, Eskişehir, Turkey Plant, Drug and Scientific Research Center, Anadolu University, Eskişehir, Turkey
Filiz Yılmaz
Affiliation:
Department of Chemistry, Anadolu University, Eskişehir, Turkey
Adil Denizli
Affiliation:
Department of Chemistry, Hacettepe University, Ankara, Turkey
Arzu Ersöz
Affiliation:
Department of Chemistry, Anadolu University, Eskişehir, Turkey
*
Corresponding author. E-mail: rsay@anadolu.edu.tr
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Abstract

The present study describes the development and use of a new bioconjugate combining targeted quantum dot labeling with an immunoperoxidase method and explores whether these bioconjugates could specifically and effectively label Cu/Zn superoxide dismutase (SOD1). The new bioconjugate is designed for the examination of samples both under fluorescent and bright-field microscopy at the same time. For this purpose chlorobis(2-2′-bipyridyl) methacryloyl tyrosine-ruthenium(II) and bis (2-2′-bipyridyl) methacryloyltyrosine-methacryloyltryptophan-ruthenium (II) photosensitive monomers and photosensitive poly(Bis (2-2′-bipyridyl)) methacryloyltyrosine-methacryloyltryptophan-ruthenium(II) were synthesized and characterized. The anti-SOD1 antibody and horseradish peroxidase (HRD) conjugated quantum dots were prepared by using this polymer. The anti-SOD1 antibody and HRD conjugated quantum dots were used in labeling and imaging of SOD1 in rat liver sections. Quantum dot particles were observed as a bright fluorescence in their specific binding locations inside the hepatocytes. The HRD-diaminobenzidine reaction product was observed as brown-colored particles at the same locations under bright-field microscopy. Structural details of the tissue sections could be examined at the same time. The conjugation protocol is simple; the bioconjugate is applicable for efficient cell labeling and can be adapted for imaging of other targets in different tissues. Also, the prepared nanobioconjugates have mechanic stability and can be used for a long period.

Type
Biological and Biomedical Applications
Copyright
Copyright © Microscopy Society of America 2012

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