Evaluation epidermal p53 immunostaining by digital image analysis.
OBJECTIVE
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.


METHODS
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).


RESULTS
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.


CONCLUSIONS
The parameters presented herein can be applied in routine analysis of immunohistochemical nuclear staining of epidermis.
Botucatu Medical School, UNESP - Univ Estadual Paulista, Department of Pathology, 18618-000, Botucatu, SP, Brazil.
Gabrielli
G
Brianezi
Eliana Maria
EM
Minicucci
Mariângela Esther Alencar
ME
Marques
Hélio Amante
HA
Miot
D001706:Biopsy; D002467:Cell Nucleus / Q000378:metabolism; D004817:Epidermis / Q000378:metabolism* / Q000473:pathology / Q000528:radiation effects; D005542:Forearm; D006801:Humans; D019044:Image Cytometry / Q000379:methods*; D007091:Image Processing, Computer-Assisted / Q000379:methods*; D007150:Immunohistochemistry / Q000379:methods*; D010781:Photography; D013472:Sunlight / Q000009:adverse effects; D016159:Tumor Suppressor Protein p53 / Q000276:immunology / Q000378:metabolism*
C495901:TP53 protein, human; D016159:Tumor Suppressor Protein p53