DEVELOPMENT OF NANOPARTICLE APPLICATIONS IN CELL IMAGING, BIOASSAY AND REACTIVE OXYGEN SPECIES DETECTION BASED ON SURFACE-ENHANCED RAMAN SPECTROSCOPY

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Description

After mid-1970, the exploration of surface-enhanced Raman scattering (SERS) has been studied by many research groups. Compared to traditional Raman spectroscopy, enormous cross section of molecules can be obtained in SERS by placing the molecules within the electromagnetic field present in metal... Show moreAfter mid-1970, the exploration of surface-enhanced Raman scattering (SERS) has been studied by many research groups. Compared to traditional Raman spectroscopy, enormous cross section of molecules can be obtained in SERS by placing the molecules within the electromagnetic field present in metal surfaces. This enhancement is due to resonance between the optical field and surface plasmon of the metal substrate. Nanoshells which are composed of a silica core and a thin gold shell can greatly enhance Raman scattering without the need to pre-aggregate the particles, due to their tunable optical property. In our study, we synthesized nanoshells as our SERS substrates for Raman tags. The stability of nanoshells coated with three different self-assembled monolayers containing poly(ethylene glycol) (PEG) molecules has been studied. Probes with Raman active PEG molecules have been delivered and imaged in macrophage cells and MCF7 cells, based on SERS technique. The benefits of this imaging technique we developed here are: 1) it is faster; 2) it requires less preparation; 3) it can provide the information of nanoshells in a semi-quantitative way in vitro. We also developed a rapid and easy-to-execute half-sandwich bioassay for the detection of low volumes (< 2 μL) of antigens on nitrocellulose membrane, based on SERS. Multiple antibodies with MW from 18.2 kDa to 170 kDa were bioconjugated to polymers and grafted to nanoshell surface to detect antigens on the membrane. SERS-based biosensors were tagged with Raman active PEGs for recognition and quantification. Here, the bioassay showed great sensitivity to very low concentration x viii of antigens and multiplexed testing have been successfully conducted on different antigens simultaneously. Moreover, SERS-based novel reactive oxygen species (ROS) sensors were designed by establishing mixed-monolayer consisting of poly(ethylene) glycol thiol (PEGSH) and either 4-nitrobenzenethiol (4-NBT) or 4-mercaptophenol (4-MP) on the surface of nanoshells. By analyzing the changes in the molecular Raman spectrum, we were able to track the production of hydroxyl radicals in low concentrations (~ 10 μM). The sensors have been delivered into the cells and did not show significant oxidative stress to the cells. Therefore, these designed sensors are very promising for tracking ROS produced by cells when they are under oxidative stress.
Ph.D. in Chemistry, July 2011 Show less

Details
Department

CHEM / Chemistry

Title

DEVELOPMENT OF NANOPARTICLE APPLICATIONS IN CELL IMAGING, BIOASSAY AND REACTIVE OXYGEN SPECIES DETECTION BASED ON SURFACE-ENHANCED RAMAN SPECTROSCOPY

Institution

Illinois Institute of Technology

Creator(s)

Yiming, Huang

Advisor(s)

Bishnoi, Sandra Whaley

Date

2011-08
2011-07

Description

After mid-1970, the exploration of surface-enhanced Raman scattering (SERS) has been studied by many research groups. Compared to traditional Raman spectroscopy, enormous cross section of molecules... Show moreAfter mid-1970, the exploration of surface-enhanced Raman scattering (SERS) has been studied by many research groups. Compared to traditional Raman spectroscopy, enormous cross section of molecules can be obtained in SERS by placing the molecules within the electromagnetic field present in metal surfaces. This enhancement is due to resonance between the optical field and surface plasmon of the metal substrate. Nanoshells which are composed of a silica core and a thin gold shell can greatly enhance Raman scattering without the need to pre-aggregate the particles, due to their tunable optical property. In our study, we synthesized nanoshells as our SERS substrates for Raman tags. The stability of nanoshells coated with three different self-assembled monolayers containing poly(ethylene glycol) (PEG) molecules has been studied. Probes with Raman active PEG molecules have been delivered and imaged in macrophage cells and MCF7 cells, based on SERS technique. The benefits of this imaging technique we developed here are: 1) it is faster; 2) it requires less preparation; 3) it can provide the information of nanoshells in a semi-quantitative way in vitro. We also developed a rapid and easy-to-execute half-sandwich bioassay for the detection of low volumes (< 2 μL) of antigens on nitrocellulose membrane, based on SERS. Multiple antibodies with MW from 18.2 kDa to 170 kDa were bioconjugated to polymers and grafted to nanoshell surface to detect antigens on the membrane. SERS-based biosensors were tagged with Raman active PEGs for recognition and quantification. Here, the bioassay showed great sensitivity to very low concentration x viii of antigens and multiplexed testing have been successfully conducted on different antigens simultaneously. Moreover, SERS-based novel reactive oxygen species (ROS) sensors were designed by establishing mixed-monolayer consisting of poly(ethylene) glycol thiol (PEGSH) and either 4-nitrobenzenethiol (4-NBT) or 4-mercaptophenol (4-MP) on the surface of nanoshells. By analyzing the changes in the molecular Raman spectrum, we were able to track the production of hydroxyl radicals in low concentrations (~ 10 μM). The sensors have been delivered into the cells and did not show significant oxidative stress to the cells. Therefore, these designed sensors are very promising for tracking ROS produced by cells when they are under oxidative stress. Show less

Type

Dissertation

File type

application/pdf

Access

Restricted Access

Rights

In Copyright
http://rightsstatements.org/page/InC/1.0/

Identifier

http://hdl.handle.net/10560/2580

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