Evaluation epidermal p53 immunostaining by digital image analysis
Article first published online: 5 JUN 2012
© 2012 John Wiley & Sons A/S
Skin Research and Technology
Volume 19, Issue 1, pages e108–e112, February 2013
How to Cite
Brianezi, G., Minicucci, E. M., Marques, M. E. A. and Miot, H. A. (2013), Evaluation epidermal p53 immunostaining by digital image analysis. Skin Research and Technology, 19: e108–e112. doi: 10.1111/j.1600-0846.2012.00616.x
- Issue published online: 7 JAN 2013
- Article first published online: 5 JUN 2012
- Manuscript Accepted: 26 APR 2012
- Manuscript Received: 25 DEC 2011
- image cytometry;
- tumor suppressor protein p53;
- image processing;
Digital techniques have been developed and validated to assess semiquantitatively immunohistochemical nuclear staining. Currently visual classification is the standard for qualitative nuclear evaluation. Analysis of pixels that represents the immunohistochemical labeling can be more sensitive, reproducible and objective than visual grading. This study compared two semiquantitative techniques of digital image analysis with three techniques of visual analysis imaging to estimate the p53 nuclear immunostaining.
Sixty-three sun-exposed forearm-skin biopsies were photographed and submitted to three visual analyses of images: the qualitative visual evaluation method (0 to 4 + ), the percentage of labeled nuclei and HSCORE. Digital image analysis was performed using ImageJ 1.45p; the density of nuclei was scored per ephitelial area (DensNU) and the pixel density was established in marked suprabasal epithelium (DensPSB).
Statistical significance was found in: the agreement and correlation among the visual estimates of evaluators, correlation among the median visual score of the evaluators, the HSCORE and the percentage of marked nuclei with the DensNU and DensPSB estimates. DensNU was strongly correlated to the percentage of p53-marked nuclei in the epidermis, and DensPSB with the HSCORE.
The parameters presented herein can be applied in routine analysis of immunohistochemical nuclear staining of epidermis.