Application of a novel high resolution widefield surface plasmon microscope in cell engineering, wound healing and development of new binding assays
Surface Plasmon (SP) microscope systems are mostly built around the prism based Krctschmann configuration. In thcsc systcms the generation of Surface Plasmons (SPs) is achieved by p-polarized light striking a metallised prism surfacc at a spccific angle and then monitoring thc intensity of the...
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my-uthm-ep.73662022-07-21T04:13:04Z Application of a novel high resolution widefield surface plasmon microscope in cell engineering, wound healing and development of new binding assays 2007 Abdul Jamil, Muhammad Mahadi QH Natural history QH201-278.5 Microscopy Surface Plasmon (SP) microscope systems are mostly built around the prism based Krctschmann configuration. In thcsc systcms the generation of Surface Plasmons (SPs) is achieved by p-polarized light striking a metallised prism surfacc at a spccific angle and then monitoring thc intensity of the reOected light. Thus in these systems. an image of the material can be obtained in terms of an intensity map. in which the intcnsity of thc image is dcpendent on the way the light couples into the SPs. The drawback of these systems is that lateral resolution relies on the ability of plasmons to propagate along the metallised layer. The lateral resolution is thus limited to a few microns. Therefore, a new microscope systcm was developed. i.e. thc Widcficld Surface Plasmon Resonance (WSPR) microscope. that is not only capable of analysing molecular interactions at high vertical resolutions. but also enables SP imaging at much higher lateral resolution than prism based systcms. The functionality of thc novel (WSPR) microscope has been investigated by imaging a scquence of binding events between micropattcrncd cxtracellular matrix proteins and their specific antibodies both in air and real-time. Using the WSPR systcm a changc in contrast was observed with each protein binding cvcnts. Images produced via the WSPR system were analyzcd and comparcd qualitatively and quantitatively. The preliminary results acquired for these binding studics between antibody/antigens dcmonstrate that the WSPR systcm capablc of resolving features down to 260nm although the theoretically proven lateral resolution of the WSPR system is -500nm. Cell surface interactions undcr two diffcrent culture conditions. i.e. HaCaTs cultured on SPR substrate with Transforming Growth Factor ~3 (TGF~3) (50ng/lII/) and without TGF~3 were also invcstigated. It was found that I-IaCaTs cultured in the presence of TGF~3 showed enhanced division and motility along with decreased cell attachmcnt as compared with cclls maintained in TGF~3 free media. It is believed that cellular signalling by TGF~3 is very important for enhancing tissue development in wound rcpair. It is confirmed that the WSPR microscope described here can be used to study sequential monomolecular layer of antibody/antigen interactions binding cvents and examination of cell surface intcrfacial intcractions at latcral scales of less than one micron without the need for traditional immunoOuorescent labelling. These rcsults have significant implications in the developmcnt of ncw brecd fast binding assays system and in enabling high resolution detailed examination of the cell surface couplings and ccll signalling proccsses involvcd in cell attachmcnt and migration. 2007 Thesis http://eprints.uthm.edu.my/7366/ http://eprints.uthm.edu.my/7366/1/24p%20MUHAMMAD%20MAHADI%20ABDUL%20JAMIL.pdf text en public phd doctoral University of Bradford School of Engineering, Design and Technology |
| institution |
Universiti Tun Hussein Onn Malaysia |
| collection |
UTHM Institutional Repository |
| language |
English |
| topic |
QH Natural history QH201-278.5 Microscopy |
| spellingShingle |
QH Natural history QH201-278.5 Microscopy Abdul Jamil, Muhammad Mahadi Application of a novel high resolution widefield surface plasmon microscope in cell engineering, wound healing and development of new binding assays |
| description |
Surface Plasmon (SP) microscope systems are mostly built around
the prism based Krctschmann configuration. In thcsc systcms the generation of
Surface Plasmons (SPs) is achieved by p-polarized light striking a metallised
prism surfacc at a spccific angle and then monitoring thc intensity of the reOected
light. Thus in these systems. an image of the material can be obtained in terms of
an intensity map. in which the intcnsity of thc image is dcpendent on the way the
light couples into the SPs. The drawback of these systems is that lateral resolution
relies on the ability of plasmons to propagate along the metallised layer. The
lateral resolution is thus limited to a few microns. Therefore, a new microscope
systcm was developed. i.e. thc Widcficld Surface Plasmon Resonance (WSPR)
microscope. that is not only capable of analysing molecular interactions at high
vertical resolutions. but also enables SP imaging at much higher lateral resolution
than prism based systcms. The functionality of thc novel (WSPR) microscope has
been investigated by imaging a scquence of binding events between
micropattcrncd cxtracellular matrix proteins and their specific antibodies both in
air and real-time. Using the WSPR systcm a changc in contrast was observed with
each protein binding cvcnts. Images produced via the WSPR system were
analyzcd and comparcd qualitatively and quantitatively. The preliminary results
acquired for these binding studics between antibody/antigens dcmonstrate that the
WSPR systcm capablc of resolving features down to 260nm although the
theoretically proven lateral resolution of the WSPR system is -500nm. Cell
surface interactions undcr two diffcrent culture conditions. i.e. HaCaTs cultured
on SPR substrate with Transforming Growth Factor ~3 (TGF~3) (50ng/lII/) and
without TGF~3 were also invcstigated. It was found that I-IaCaTs cultured in the
presence of TGF~3 showed enhanced division and motility along with decreased
cell attachmcnt as compared with cclls maintained in TGF~3 free media. It is
believed that cellular signalling by TGF~3 is very important for enhancing tissue
development in wound rcpair. It is confirmed that the WSPR microscope
described here can be used to study sequential monomolecular layer of
antibody/antigen interactions binding cvents and examination of cell surface
intcrfacial intcractions at latcral scales of less than one micron without the need
for traditional immunoOuorescent labelling. These rcsults have significant
implications in the developmcnt of ncw brecd fast binding assays system and in
enabling high resolution detailed examination of the cell surface couplings and
ccll signalling proccsses involvcd in cell attachmcnt and migration. |
| format |
Thesis |
| qualification_name |
Doctor of Philosophy (PhD.) |
| qualification_level |
Doctorate |
| author |
Abdul Jamil, Muhammad Mahadi |
| author_facet |
Abdul Jamil, Muhammad Mahadi |
| author_sort |
Abdul Jamil, Muhammad Mahadi |
| title |
Application of a novel high resolution widefield surface plasmon microscope in cell engineering, wound healing and development of new binding assays |
| title_short |
Application of a novel high resolution widefield surface plasmon microscope in cell engineering, wound healing and development of new binding assays |
| title_full |
Application of a novel high resolution widefield surface plasmon microscope in cell engineering, wound healing and development of new binding assays |
| title_fullStr |
Application of a novel high resolution widefield surface plasmon microscope in cell engineering, wound healing and development of new binding assays |
| title_full_unstemmed |
Application of a novel high resolution widefield surface plasmon microscope in cell engineering, wound healing and development of new binding assays |
| title_sort |
application of a novel high resolution widefield surface plasmon microscope in cell engineering, wound healing and development of new binding assays |
| granting_institution |
University of Bradford |
| granting_department |
School of Engineering, Design and Technology |
| publishDate |
2007 |
| url |
http://eprints.uthm.edu.my/7366/1/24p%20MUHAMMAD%20MAHADI%20ABDUL%20JAMIL.pdf |
| _version_ |
1747831141117722624 |