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

@article{citeulike:2369260, abstract = {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 A1 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 A1 receptor quantification was found (r = 0.98, P < 0.001, and 95\% CI = 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 immunohistochemical staining area (occupied by a specific antigen) in small sample tissues that do not fill the entire image field of the microscope.}, author = {Leal, Sandra and Diniz, Carmen and Sa, Carlos and Goncalves, Jorge and Soares, Ana S. and Rocha-Pereira, Carolina and Fresco, Paula }, citeulike-article-id = {2369260}, doi = {http://dx.doi.org/10.1016/j.ab.2006.07.017}, journal = {Analytical Biochemistry}, keywords = {dab, image-analysis, immunohistochemistry, microscopy}, month = {October}, number = {1}, pages = {137--143}, posted-at = {2008-02-13 15:07:27}, priority = {3}, title = {Semiautomated computer-assisted image analysis to quantify 3,3'-diaminobenzidine tetrahydrochloride-immunostained small tissues}, url = {http://dx.doi.org/10.1016/j.ab.2006.07.017}, volume = {357}, year = {2006} }

TY - JOUR ID - citeulike:2369260 L3 - citeulike-article-id:2369260 TI - Semiautomated computer-assisted image analysis to quantify 3,3'-diaminobenzidine tetrahydrochloride-immunostained small tissues JF - Analytical Biochemistry VL - 357 IS - 1 SP - 137 EP - 143 N2 - 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 A1 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 A1 receptor quantification was found (r = 0.98, P < 0.001, and 95% CI = 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 immunohistochemical staining area (occupied by a specific antigen) in small sample tissues that do not fill the entire image field of the microscope. KW - dab KW - image-analysis KW - immunohistochemistry KW - microscopy AU - Leal, Sandra AU - Diniz, Carmen AU - Sa, Carlos AU - Goncalves, Jorge AU - Soares, Ana AU - Rocha-Pereira, Carolina AU - Fresco, Paula PY - 2006/10/01/ UR - http://dx.doi.org/10.1016/j.ab.2006.07.017 ER -