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| Home > Publications database > Detectability of radiation induced H2O2 with lowconcentrated fluorophores NucPE1 and PY1-ME |
| Home > Publications database > Detectability of radiation induced H2O2 with lowconcentrated fluorophores NucPE1 and PY1-ME |
| Bachelor Thesis | DKFZ-2021-02543 |
2021
Abstract: A new method to quantify the amount of DNA damage due to irradiation is currentlydeveloped for applications in radiation therapy research. It quantifies the amount ofDNA damage by measuring the amount of produced hydrogen peroxide H2O2. This willbe measured inside cells with the fluorophores Nuclear Peroxy Emerald 1(NucPE1) andPeroxy Yellow-1 Methyl Ester (PY1-ME).In this thesis the number of molecules of each dye that localise inside a cell of the cell lineH460 was analysed. This resulted in an amount of (1.4±0.3)108 moleculescell for NucPE1 and(1 ± 3)109 moleculescell for PY1-ME.In addition a function to calibrate a number n of H460 cells to a concentration c was foundto be c = f ∗ n + bg. 100 µL of a solution with concentration c of the fluorophore producea signal of the same magnitude as n cells. The parameters of this function were found tobe fNucP E1 = (2.4 ± 0.5)10−6 µMcell and bgNucP E1 = (0 ± 1.8) µM for NucPE1. For PY1-MEthe parameters resulted in fP Y 1−ME = (2 ± 5)10−5 µMcell and bgP Y 1−ME = (3 ± 82) µM.Moreover, it was possible to detect the increase of fluorescence signal with increasing dosefor a number of molecules like there are in 5000 H460 cells. This was realised with a platereader and a photon counting system.The results confirm the possibilty to developing a chemical dosimeter for in cell measurements based on one of these fluorophores.
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