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Entwicklung und Einsatz der Immun-SERS-Mikroskopie zur Gewebe-basierten Tumordiagnostik

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https://osnadocs.ub.uni-osnabrueck.de/handle/urn:nbn:de:gbv:700-2013090911535
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dc.contributor.advisorProf. Dr. Sebastian Schlücker
dc.creatorSalehi, Mohammad
dc.date.accessioned2013-09-09T13:49:30Z
dc.date.available2013-09-09T13:49:30Z
dc.date.issued2013-09-09T13:49:30Z
dc.identifier.urihttps://osnadocs.ub.uni-osnabrueck.de/handle/urn:nbn:de:gbv:700-2013090911535-
dc.description.abstractSurface-enhanced Raman scattering (SERS) microscopy is a novel method of optical imaging for the localization and quantification of target molecules in cells and tissue specimens. The major advantages of SERS over fluorescence are quantification and spectral multiplexing due to the small line width of vibrational Raman bands. The position of the plasmon band of both hollow gold/silver nanoshells and silica-encapsulated gold nanoclusters can be tuned for maximum SERS enhancement upon red laser excitation, which is optimal for minimizing the disturbing autofluorescence of tissue. In this work, silica-encapsulated and non-encapsulated SERS particles were used for the localization of target proteins in prostate tissue specimens. Two different biofunctionalization methods were established for each type of SERS particles. The cross-linking method based on s-NHS/EDC chemistry was modified for covalently conjugating proteins to hollow gold/silver nanoshells and gold nanostars in order to minimize the aggregation of SERS nanoparticles during and after cross-linking. As an alternative to covalent conjugation chemistry, the noncovalent binding of antibodies to the SERS particles via an adapter protein (protein A/G) was established. The influence of several factors that determine the quality of results obtained by SERS imaging, such as the number of immuno-SERS conjugates, incubation time, antigen retrieval and blocking buffer, were investigated. Rapid SERS microscopy with 30 msec acquisition time per pixel was enabled by using silica-encapsulated gold nanoclusters for the localization of p63 proteins on prostate tissue specimens from healthy donors. Two-color SERS experiments for the parallel localization of PSA and p63 were performed with silica-encapsulated and non-encapsulated nanoshells. The quality of the results depends less on the nature of the surface chemistry of the nanoparticles (with or without silica encapsulation), but more on the blocking buffer and the antigen retrieval method. Silica-encapsulated gold nanoclusters were also used for the simultaneous quantification of three cytokines (IL1, IL8 and TNF- α) in a SERS-based sandwich immunoassay with a detection limit of ca. 0.3 pM. Keywords: Raman, SERS microscopy, biocompatibility of nanoparticles, cross-linking, antigen unmasking methods, antigen detection, immunohistochemistry, immunoassay.eng
dc.subjectSERSger
dc.subjectRamanger
dc.subjectNanopartikelger
dc.subjectImmunhistochemieger
dc.subjectCrosslinkingger
dc.subjectImmunglobulinger
dc.subjectImmunoassayger
dc.subject.ddc530 - Physik
dc.subject.ddc570 - Biowissenschaften; Biologie
dc.subject.ddc540 - Chemie
dc.subject.ddc610 - Medizin und Gesundheit
dc.titleEntwicklung und Einsatz der Immun-SERS-Mikroskopie zur Gewebe-basierten Tumordiagnostikger
dc.typeDissertation oder Habilitation [doctoralThesis]-
thesis.locationOsnabrück-
thesis.institutionUniversität-
thesis.typeDissertation [thesis.doctoral]-
thesis.date2013-08-05-
dc.contributor.refereePD. Dr. Jens Packeisen
dc.subject.bk33.18 - Optik
dc.subject.bk35.10 - Physikalische Chemie: Allgemeines
dc.subject.bk42.03 - Methoden und Techniken der Biologie
dc.subject.bk44.45 - Immunologie
dc.subject.bk44.47 - Pathologie
dc.subject.bk44.81 - Onkologie
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