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Evaluation epidermal p53 immunostaining by digital image analysis

Authors

  • Gabrielli Brianezi,

    1. Botucatu Medical School, UNESP - Univ Estadual Paulista, Department of Pathology, Botucatu, SP, Brazil
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  • Eliana Maria Minicucci,

    1. Botucatu Medical School, UNESP - Univ Estadual Paulista, Department of Dermatology and Radiotherapy, Botucatu, SP, Brazil
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  • Mariângela Esther Alencar Marques,

    1. Botucatu Medical School, UNESP - Univ Estadual Paulista, Department of Pathology, Botucatu, SP, Brazil
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  • Hélio Amante Miot

    Corresponding author
    1. Botucatu Medical School, UNESP - Univ Estadual Paulista, Department of Dermatology and Radiotherapy, Botucatu, SP, Brazil
    • Botucatu Medical School, UNESP - Univ Estadual Paulista, Department of Pathology, Botucatu, SP, Brazil
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Address:

Hélio Amante Miot

Departamento de Dermatologia da Faculdade de Medicina da Unesp

S/N.Campus Universitário de Rubião Jr.

18618-000

Botucatu SP

Brazil

Tel./Fax: +55 14 38824922

e-mail: heliomiot@fmb.unesp.br

Abstract

Background/purpose

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.

Conclusion

The parameters presented herein can be applied in routine analysis of immunohistochemical nuclear staining of epidermis.

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