Nanoparticle-based improvement of the FISH and light microscopy techniques for contrast enhancement in environmental microbiology. | |
| Author | Bonduush, Ichinkhorloo. |
| Call Number | AIT Thesis no.EV-07-15 |
| Subject(s) | Nanoparticles Fluorescence in situ hybridization |
| Note | A thesis submitted in partial fulfillment of the requirements for the degree of Master of Science in Environmental Engineering and Management Inter-University Program on Environmental Toxicology, Technology and Management. |
| Publisher | Asian Institute of Technology |
| Series Statement | Thesis ; no. EV-07-15 |
| Abstract | Luminescent quantum dots (QDs)-semiconductor nanocrystals are a promising alternative to organic dyes for fluorescence-based applications. The photostability and narrow emission spectra of non-organic Quantum Dot fluorophores (QDs) make them desirable candidates for Fluorescent In Situ Hybridization (FISH). In this study we demonstrated the first application of the luminescence ZnS:Mn2+QDs for development of conventional FISH protocol and a continuation of the prior research on the application of chitosan coated and glutamate coated gold nanoparticles for biological image enhancement of LM. To the best of our knowledge, this is the first report FISH technique improvement by using ZnS:Mn2+QD-labeled DNA probes to detect different Bacteria and Archaea. FISH analysis using quantum dot probes requires the QDs be soluble and stable in solution which used in hybridization procedure such as blocking, hybridization and washing solutions, and that the dot surface be efficiently derivatized with DNA probes. In this regard we have developed the surface modification procedures of Chitosan coated and noncoated of ZnS:Mn2+QD to obtain successful conjugation between the QDs and metal-thiol labeled DNA probes. In order hand our probe is bigger than pores of cell membranes. Therefore we also developed additional treatment procedure of fixed bacterial cells. Our experiments have shown that by QD-FISH technique selected microorganisms can be seen brightly fluorescing without the use of any fluorescent dyes. One of major advantages is non-toxicity of ZnS:Mn2+QD in comparison to other known QDs, e.g. CdSe and core shell CdSc/Zn. Another purpose of this thesis was to investigate contrast enhancement of LM using Chitosan coated and glutamate-coated colloidal gold nanoparticle (GNPs) solutions. According to the application of both types of GNP we found that chitosan coated GNPs substantially enhanced the contrast of LM as a result of attaching to the surface of microorganisms. |
| Year | 2007 |
| Corresponding Series Added Entry | Asian Institute of Technology. Thesis ; no. EV-07-15 |
| Type | Thesis |
| School | School of Environment, Resources, and Development (SERD) |
| Department | Department of Energy and Climate Change (Former title: Department of Energy, Environment, and Climate Change (DEECC)) |
| Academic Program/FoS | Environmental Engineering and Management (EV) |
| Chairperson(s) | Shipin, Oleg; |
| Examination Committee(s) | Dutta, Joydeep;Skorn Mongkolsuk; |
| Scholarship Donor(s) | JAPAN; |
| Degree | Thesis (M.Sc.) - Asian Institute of Technology - Chulabhorn Research Institute - Mahidol University, 2007 |