Assessment of cyclobutane pyrimidine dimers by digital photography in human skin

作者：Ten Berge Onno^*; van Velsen Sara G A; Giovannone Barbara; Bruijnzeel Koomen Carla A F M; Knol Edward F; Guikers Kees; van Weelden Huib

来源：Journal of Immunological Methods, 2011, 373(1-2): 240-246.

DOI：10.1016/j.jim.2011.07.014

• DNA
• UVB
• DNA damage repair
• Assessment
• Thymine dimer

UV-mediated DNA damage and repair are important mechanisms in research on UV-induced carcinogenesis. UV-induced DNA-damage and repair can be determined by immunohistochemical staining of photoproduct positive nuclei of keratinocytes in the epidermis. We developed a new method of analysing and quantifying thymine dimer (TT-CPD) positive cells in the epidermis.
Normal skin of healthy controls was exposed to UVB ex vivo and in vivo. Skin samples were immunohistochemically stained for TT-CPDs. Digital images of the epidermis were quantified for TT-CPDs both visually and digitally.
There was a UVB-dose dependent induction of TT-CPDs present in the ex vivo UVB-irradiated skin samples. The linear measurement range of the digital quantification was increased compared to the manual counting. The average 24-hour repair rate of the initiated TT-CPDs elicited by the UVB irradiation at T = 0 of the 8 HCs showed a 34% decrease of TT-CPD photoproducts by the manual counting method and a 51% decrease determined by digital counting.
The digital quantification method improves immunohistochemical quantification of DNA photo damage. It is more sensitive in measuring the extent of DNA-damage per nucleus.

• 出版日期2011-10-28