guest ::
| Home > Publications database > Detection of Radiation Induced H2O2 in H460Lung Cancer Cells Using Novel NucPE1 Dye |
| Home > Publications database > Detection of Radiation Induced H2O2 in H460Lung Cancer Cells Using Novel NucPE1 Dye |
| Bachelor Thesis | DKFZ-2020-02655 |
2020
Abstract: The tumor response after irradiation depends largely on the applied dose and thereby the energy transferred to the cells. A possible newway to measure this deposited energy was found with the development of thenuclear-localized fluorescent hydrogen peroxide (H2O2) probe Nuclear PeroxyEmerald 1 (NucPE1). It is capable of detecting H2O2 inside the nucleus, areactive oxygen species (ROS) that is often induced by radiation and playsan important role in the production of indirect DNA damage. By measuringthe fluorescent signal of the nuclei of cells incubated with NucPE1 and treatedwith ionizing radiation, it can be possible to gain valuable insight into thisdose-response relationship.In this thesis, some of the general characteristics of NucPE1 were examined,including its stability in solution and its interaction with artificially addedH2O2. The time consistency of the fluorescent NucPE1 signal from H460 lungcancer cells was measured, and different protocols were investigated that couldpotentially increase its lifetime. A protocol to freeze and fix the NucPE1 stainedcells was developed, which allowed measurement of a rather stable signal for upto 45min after irradiation. The fluorescent intensities of H460 cells irradiatedwith different doses were measured using confocal microscopy and the nuclei ofstained cells were extracted and analyzed using flow cytometry (FACS). ThisFACS analysis led to the most promising results that almost fit the expectationof a linear-quadratic NucPE1-dose-response. With this, a large step towardsthe usage of NucPE1 as a chemical dosimeter has been achieved.
|
The record appears in these collections: |