The quantitation and kinetics of unscheduled (repair) DNA synthesis in ultraviolet-irradiated human skin by automated image analysis

Summary Grain counting by eye is a tedious and time‐consuming technique but one with great potential in cell kinetics and for the study of DNA excision repair activity (unscheduled DNA synthesis or UDS). We have been investigating the levels of UDS in human skin sections exposed in situ to ultraviol...

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Bibliographic Details
Published in:British journal of dermatology (1951) 1996-10, Vol.135 (4), p.516-522
Main Authors: CHADWICK, C.A., DIGGLE, S.P., YOUNG, A.R., POTTEN, C.S.
Format: Article
Language:English
Subjects:
Autoradiography - methods
Biological and medical sciences
DNA - biosynthesis
DNA Repair - radiation effects
Humans
Image Processing, Computer-Assisted - methods
Investigative techniques, diagnostic techniques (general aspects)
Medical sciences
Otorhinolaryngology. Stomatology. Orbit
Radiodiagnosis. Nmr imagery. Nmr spectrometry
Skin - metabolism
Skin - radiation effects
Thymidine
Tritium
Ultraviolet Rays
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Summary:Summary Grain counting by eye is a tedious and time‐consuming technique but one with great potential in cell kinetics and for the study of DNA excision repair activity (unscheduled DNA synthesis or UDS). We have been investigating the levels of UDS in human skin sections exposed in situ to ultraviolet radiation using a short‐term incubation in tritiated thymidine and autoradiography and the decline in UDS levels with time (repair Kinetics). We have adapted an automated image analysis system automatically to assess the number of grains over epidermal cell nuclei in autoradiographs of sections of epidermis. An excellent correlation was observed between visual counting and machine measurement of the area (in pixels) occupied by silver grains. The levels of UDS declined with time as lesions are progressively repaired. The half time (± standard deviation) for the reduction in UDS is 7.25± 0.18 h. The grain counts can be significantly increased by increasing the autoradio‐graphic exposure, by increasing the concentration of tritiated thymidine and by increasing the incubation time.
ISSN:0007-0963
1365-2133
DOI:10.1046/j.1365-2133.1996.d01-1032.x