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@MASTERSTHESIS{Voigt:166147,
author = {J. Voigt$^*$},
title = {{D}etection of {R}adiation {I}nduced {H}2{O}2 in
{H}460{L}ung {C}ancer {C}ells {U}sing {N}ovel {N}uc{PE}1
{D}ye},
school = {Universität Heidelberg},
type = {Bachelorarbeit},
reportid = {DKFZ-2020-02655},
year = {2020},
note = {Bachelorarbeit, Universität Heidelberg, 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.},
cin = {E041},
cid = {I:(DE-He78)E041-20160331},
pnm = {315 - Imaging and radiooncology (POF3-315)},
pid = {G:(DE-HGF)POF3-315},
typ = {PUB:(DE-HGF)2},
url = {https://inrepo02.dkfz.de/record/166147},
}