Immunohistochemical determination of nuclear antigens by colour image analysis: application for labelling index, estrogen and progesterone receptor status in breast cancer.
The immunohistochemical evaluation of 5-bromo-2'-deoxyuridine (BrdU) labelling index, estrogen (ER) and progesterone receptor (PR) status was carried out on the automated computer-assisted image analysis station BIOCOM 500. Special software has been developed to measure nuclear antigens using the immunoperoxidase method with the Harris hematoxylin counterstain. The analysis was based on the different light adsorption spectra of the chromogen diaminobenzidine and Harris's hematoxylin coloration when exposed to light of differing wavelengths. The results obtained by image analysis were compared to previously validated methods. The thymidine labelling index performed by manual procedures and BrdU incorporation performed by image analysis were comparable (linear correlation coefficient r = 0.76, P < 0.001). Comparison of image analysis and dextran coated charcoal assay for ER and PR content revealed excellent sensitivities and specificities (linear correlation coefficient r = 0.87, P < 0.001 for ER and r = 0.93, P < 0.001 for PR). These data suggest that automated image analysis offers a reliable and reproducible procedure for measuring nuclear antigens.
Rostagno
P
P
Laboratoire de cytométrie, Centre Antoine Lacassagne, Nice, France.
Birtwisle
I
I
Ettore
F
F
Courdi
A
A
Gioanni
J
J
Namer
M
M
Caldani
C
C
D000951:Antigens, Neoplasm / Q000032:analysis*; D001943:Breast Neoplasms / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism; D001973:Bromodeoxyuridine; D003116:Color; D006801:Humans; D007091:Image Processing, Computer-Assisted; D007124:Immunoenzyme Techniques; D009687:Nuclear Proteins / Q000032:analysis*; D011960:Receptors, Estrogen / Q000032:analysis*; D011980:Receptors, Progesterone / Q000032:analysis*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013936:Thymidine / Q000378:metabolism
D000951:Antigens, Neoplasm; D009687:Nuclear Proteins; D011960:Receptors, Estrogen; D011980:Receptors, Progesterone; D001973:Bromodeoxyuridine; D013936:Thymidine