Quantitative analysis of DNA-damage response factors after sequential ion microirradiation.
Several proteins are known to form foci at DNA sites damaged by ionizing radiation. We study DNA damage response by immunofluorescence microscopy after microirradiation of cells with energetic ions. By using microirradiation, it is possible to irradiate different regions on a single dish at different time-points and to differentiate between cells irradiated earlier and later. This allows to directly compare immunofluorescence intensities in both subsets of cells with little systematic error because both subsets are cultivated and stained under identical conditions. In addition, by using irradiation patterns such as crossing lines, it is possible to irradiate individual cells twice and to differentiate between immunofluorescence signals resulting from the cellular response to the earlier and to the later irradiation event. Here, we describe the quantitative evaluation of immunofluorescence intensities after sequential irradiation.
Greubel
Christoph
C
LRT2, Universität der Bundeswehr München, 85579 Neubiberg, Germany. christoph.greubel@unibw.de
Hable
Volker
V
Drexler
Guido A
GA
Hauptner
Andreas
A
Dietzel
Steffen
S
Strickfaden
Hilmar
H
Baur
Iris
I
Krücken
Reiner
R
Cremer
Thomas
T
Friedl
Anna A
AA
Dollinger
Günther
G
D004249:DNA Damage / Q000502:physiology*; D004273:DNA, Neoplasm / Q000235:genetics* / Q000528:radiation effects* / Q000648:ultrastructure; D004307:Dose-Response Relationship, Radiation; D006367:HeLa Cells; D006801:Humans; D007090:Image Interpretation, Computer-Assisted / Q000379:methods*; D007477:Ions; D008856:Microscopy, Fluorescence / Q000379:methods*; D011829:Radiation Dosage; D011874:Radiometry / Q000379:methods*
D004273:DNA, Neoplasm; D007477:Ions