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Interactions between silver nanoparticles and fluorescent phytochromes from Deinococcus radiodurans

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Authors
Nuuttila, Lauri
Date
2017
Discipline
FysiikkaPhysics

 
Poikkitieteelliset sovellukset ovat viime aikoina yleistyneet tieteellisessä tutkimuksessa. Tämä näkyy hyvin esimerkiksi elävän kudoksen kuvantamisen kehittymisessä, jota varten etsitään jatkuvasti parempia biologisia leimoja fysiikan ja kemian keinoin. Tässä tutkielmassa on kehitetty ensimmäinen lähi-infrapuna-alueen fluoresenssileima, joka hyödyntää hopeananopartikkeleja ja fytokromeja. Kompleksi on syntetisoitu itse ja sen rakenne on todennettu optisen spektroskopian, elektroforeesin ja elektronimikroskopian keinoin. Lupaavat fluoresenssiominaisuudet saatiin käyttämällä monomeerisen Deinococcus radiodurans -fytokromin kromoforia sitovaa domeenia (CBDmon). Hopeapartikkelien läsnäollessa CBDmonin kvanttisaanto kasvoi 20 % ja viritetyn tilan elinaika laski 30 %. Syntetisoidun kompleksin kirkkaus verrattuna CBDmonin kirkkauteen oli 470+-40% 390 nm:n ja 134+-11% 630 nm:n virityksellä. Näitä tuloksia voidaan todennäköisesti vielä kasvattaa, koska kompleksin rakennetta on mahdollista muokata monella eri tavalla. Siksi voidaan todeta, että kehitetty menetelmä on lupaava keino lähi-infrapunaalueen leimojen suunnitteluun. ...
 
Applications using a cross-disciplinary approach are a common phenomenon in recent scientific research. A good example is the field of living tissue imaging where better biological labels are constantly searched with the help of physics and chemistry. In this Thesis, a near-infrared template combining silver nanoparticles and phytochromes was synthesized. The formation of the complex was confirmed by using optical spectroscopy, electrophoresis and electron microscopy. Promising fluorescent properties were obtained using monomeric chromophore binding domain (CBDmon) of a phytochrome from Deinococcus radiodurans. In the presence of silver nanoparticles, CBDmon showed 20% increased quantum yield and 30% decreased exited state lifetime. The brightness of the synthesized complex was 470+-40% at 390 nm excitation wavelength and 134+-11% at 630 nm relative to the brightness of pure CBDmon. It is likely that these promising properties could be improved by further engineering using variety of methods. Thus, the results indicate a new way to design applicable NIR-labels. ...
 
Keywords
plasmonit kirkkaus CBDmon absorptio fluoroforit kvanttisaanto säteily elinaika plasmon brightness imaging absorption fluorophore quantum yield radiation lifetime kuvantaminen fluoresenssimikroskopia kemialliset yhdisteet fluoresenssi
URI

http://urn.fi/URN:NBN:fi:jyu-201709153737

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