Semiautomated computer-assisted image analysis to quantify 3,3 '-diaminobenzidine tetrahydrochloride-immuno stained small tissues

• Type: Article in International Scientific Journal
• Date: 2006

Authors

Sandra Leal (Author) Other; Carmen Diniz (Author) FFUP; Carlos Sa (Author) FEUP; Jorge Goncalves (Author) FFUP; Ana Sofia Soares (Author) Other; Carolina Rocha Pereira (Author) Other; Paula Fresco (Author) FFUP

Journal

Title: Analytical Biochemistry

Vol. 357

Pages: 137-143

ISSN: 0003-2697

Publisher: Elsevier

Indexing

ISI Web of Knowledge - 24 Citations

Scopus - 24 Citations

Scientific classification

FOS: Natural sciences > Chemical sciences

Other information

Authenticus ID: P-004-GP5

DOI: 10.1016/j.ab.2006.07.017

Abstract (EN): This work aimed to develop a technique to measure stained areas in images from sample tissue sections, namely when the structure of interest does not fill the entire image field of the microscope. We propose a semiautomated computer-assisted image analysis (SACAIA) method in which brightfield color images of 3,3'-diaminobenzidene tetrahydrochloride (DAB)-stained antigens are converted to their blue component and boundaries are delineated to extract the object of interest. The number of pixels of a defined color (elicited by DAB) is counted and used to measure the stained area relative to the total area of the tissue under study. The percentages of area stained with adenosine A(1) receptor were 40.76 +/- 2.08 and 42.44 +/- 2.26% for manual analysis and SACAIA, respectively (P = 0.582). A strong linear correlation of A(1) receptor quantification was found (r = 0.98, P < 0.001, and 95% Cl = 0.97 to 0.99 for manual method; r = 0.99, P < 0.001, and 95% CI = 0.98 to 0.99 for SACAIA method). The extent to which misclassification affected staining quantification was evaluated by Bland-Altman analysis, indicating that this method can be applied accurately to quantify the immumohistochemical staining area (occupied by a specific antigen) in small sample tissues that do not fill the entire image field of the microscope.

Language: English

Type (Professor's evaluation): Scientific

No. of pages: 7