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Conjugated polythiophene probes target lysosome-related acidic vacuoles in cultured primary cells
Linköpings universitet, Institutionen för fysik, kemi och biologi, Biomolekylär och Organisk Elektronik. Linköpings universitet, Tekniska högskolan.
Linköpings universitet, Institutionen för fysik, kemi och biologi, Organisk Kemi. Linköpings universitet, Tekniska högskolan.ORCID-id: 0000-0002-5582-140X
Linköpings universitet, Institutionen för klinisk och experimentell medicin, Klinisk kemi. Linköpings universitet, Hälsouniversitetet.
Linköpings universitet, Institutionen för klinisk och experimentell medicin, Klinisk kemi. Linköpings universitet, Hälsouniversitetet.
Vise andre og tillknytning
2007 (engelsk)Inngår i: Molecular and Cellular Probes, ISSN 0890-8508, Vol. 21, nr 5-6, s. 329-337Artikkel i tidsskrift (Fagfellevurdert) Published
Abstract [en]

Conformation-sensitive optical probes for studying biological processes and structures are of great interest. The present work shows for the first time that conjugated polyelectrolyte (CPE) probes can be used for specific targeting of chromatin, nuclear and cytoplasmatic vesicles, and cytoskeletal components in a complex system of cultured cells. One of the probes could also be used for vital staining of live cells. When bound to different entities, the polythiophene derivative probes emitted light with different colors due to the unique spectral properties of these conformation sensitive probes. The physical pre-requisites for binding could also be exploited for characterization of the target. Unexpectedly, lysosome-related acidic vacuoles were targeted in cultured primary cells by both anionic, cationic, and zwitter-ionic polythiophene derivatives. Pre-treatment with Bafilomycin A1, a specific inhibitor of vacuolar-type H+-ATPase, caused redistribution of the staining. The targeting of lysosome-related acidic vesicles could not be demonstrated in transformed cells (melanoma, neuroblastoma, and prostate cancer cell lines), indicating a difference in the localization, structure, accessibility, or quantity of the target in cultured normal cells as compared with the malignant cell lines. The chemical nature of the conjugated polyelectrolyte complex in the cytoplasmatic vacuoles remains elusive.

sted, utgiver, år, opplag, sider
2007. Vol. 21, nr 5-6, s. 329-337
Emneord [en]
Conjugated polyelectrolyte; Polythiophene; Fibroblast; Lysosome; Cancer
HSV kategori
Identifikatorer
URN: urn:nbn:se:liu:diva-12727DOI: 10.1016/j.mcp.2007.04.005OAI: oai:DiVA.org:liu-12727DiVA, id: diva2:16929
Tilgjengelig fra: 2007-12-07 Laget: 2007-12-07 Sist oppdatert: 2014-04-08
Inngår i avhandling
1. Biological Sensing and DNA Templated Electronics Using Conjugated Polymers
Åpne denne publikasjonen i ny fane eller vindu >>Biological Sensing and DNA Templated Electronics Using Conjugated Polymers
2007 (engelsk)Doktoravhandling, med artikler (Annet vitenskapelig)
Abstract [en]

Conjugated polymers have been found useful in a wide range of applications such as solar cells, sensor elements and printed electronics, due to their optical and electronic properties. Functionalization with charged side chains has enabled water solubility, resulting in an enhanced interaction with biomolecules. This thesis focus on the emerging research fields, where these conjugated polyelectrolytes (CPEs) are combined with biomolecules for biological sensing and DNA nanowire assembling.

CPEs have shown large potential in biomolecular detection where the optical read out is due to the geometrical alternation in the backbone and aggregation state. This thesis focused on transferring the biomolecular detection to a surface of CPEs. The characterization of the CPE layer show that a hydrogel can be formed, and how the layer can undergo geometrical changes upon external stimulus such as pH change. A selective sensor surface can be created by imprinting ssDNA or an antibody in the CPE layer. The discrimination for complementary DNA hybridization and specific antibody interaction can be monitored by surface plasmon resonance or quartz crystal microbalance. We have also taken the step out from the controlled test tube experiments to the complex environment of the cell showing the potential for staining of compartments and structures in live and fixed cell. Depending on the conditions and CPE used, cell nuclei, acidic vesicles and cytoskeleton structure can be visualized. Furthermore, the live staining shows no sign of toxic effect on cultured fibroblasts.

CPEs can also be a valuable element when assembling electronics in the true nano regime. I have used DNA as building template due to its attractive size features, with a width of around 2 nm and a length scale in the µm regime, and the inbuilt base-paring recognition elements. This thesis shows how DNA can be decorated with CPEs and stretched on surfaces into a model for aligned semiconducting nanowire geometries. Not only making the template structures is of importance, but also how to place them on the correct surface position, i.e. on electrodes. Strategies for positioning DNA nanowires using transfer printing and surface energy patterning methods have therefore been developed in the thesis. The stretched DNA decorated with CPE also offers a way to further study the molecular binding interaction between the two molecules. Single molecular spectroscopy in combination with polarization has given information of the variation of the CPE binding along a DNA chain.

sted, utgiver, år, opplag, sider
Institutionen för fysik, kemi och biologi, 2007
Serie
Linköping Studies in Science and Technology. Dissertations, ISSN 0345-7524 ; 1154
Emneord
conjugated polymer, conjugated polyelectrolyte, DNA, sensor, nanowire
HSV kategori
Identifikatorer
urn:nbn:se:liu:diva-10180 (URN)978-91-85895-17-5 (ISBN)
Disputas
2007-12-14, Planck, Fysikhuset, Campus Valla, Linköpings Universitet, Linköping, 10:15 (engelsk)
Opponent
Veileder
Tilgjengelig fra: 2007-12-07 Laget: 2007-12-07 Sist oppdatert: 2009-04-23

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