QUANTUM DOT BASED BIOSENSING
Özet
Semiconductor nanocrytals also known as quantum dots (QD) with high photoluminesce quantum yield (PLQY), size tunability and favorable optical characteristics occupy a significant area in display technology, solar energy conversion and bioapplications. Size tuning feature of QDs allows emission wavelength ranging from ultraviolet to infrared spectral region. In literature, QD based studies have been performed in visible spectral range by employing mostly cadmium, being a toxic heavy metal. Recently, the search for less toxic alternatives revealed the cadmium free compounds, particularly InP. Cadmium free semiconductor nanocrytals’ potential to be used as fluorescent probes in biodetection and biolabeling area has been proved over the past decades.
Pathogens threaten life particularly via water sources like rivers, reservoirs and groundwater. Increasing demand for managing the ‘contamination of drinkable water by pathogenic bacteria’ problem needs a broad perspective about pathogens and their membrane characteristics which are integral part of microorganism detection platforms. Bacteria are categorized mainly upon their membrane properties which are gram negative and gram positive. Extra wall called as peptidoglycan layer in gram positive bacteria makes them more resistant to external forces. Gram negative bacteria with wavy wall is relatively more prone to their environment. One of the most known pathogenic bacteria, E. Coli, have damaged and destroyed many lives throughout the world. High growth rate enables this microorganism to spread around large areas in short time. Therefore, accurate and definite detection of this bacteria in water is crucial.
The main frame of this research depends on QD based biodetection of bacteria. First of all, organic based QDs (50% PLQY) containing triocytlyphosphine-sulfur ligand were synthesized and via successful phase transfer, aqueous QDs with 20% PLQY were achieved. Although surface is damaged during ligand exchange procedure, aqueous QDs with high PLQY were obtained. SiO2 was covered with QDs thanks to the attraction between their NH2 group and carboxylic ends, respectively. In the final step, this hybrid structure was covered with SiO2 and silica coated QDs (SCQD) were formed. In order to utilize SCQDs in bacteria detection, fluorescent agents were embeded in polymeric films which were formed by spin coating. As a result, SCQD facilitates the attachment of negatively charged bacteria onto the surface of the films.
Appropriately grown DH5 alpha (E. Coli strain) expressing green fluorescent protein (GFP) was used as pathogen in the detection part. SCQD thin films were treated with water containing E.Coli DH5 alpha. Positively charged SCQD attracted negatively charged bacteria and the conjugation between them was analysed with time resolved spectroscopy and monitored with fluorescence microscope. Thus, usage of QDs as biosensor in pathogen detection could provide an insight in the future studies.