TY - JOUR
T1 - Accurate quantitation of Ki67-positive proliferating hepatocytes in rabbit liver by a multicolor immunohistochemical (IHC) approach analyzed with automated tissue and cell segmentation software
AU - van der Loos, Chris M.
AU - de Boer, Onno J.
AU - Mackaaij, Claire
AU - Hoekstra, Lisette T.
AU - van Gulik, Thomas M.
AU - Verheij, Joanne
PY - 2013/1
Y1 - 2013/1
N2 - Determination of hepatocyte proliferation activity is hampered by the presence of Ki67-positive non-parenchymal cells. We validated a multicolor immunohistochemical (IHC) approach using multispectral tissue and cell segmentation software. Portal vein branches to the cranial liver lobes of 10 rabbits were embolized, leading to atrophy of the cranial lobes and hyperplasia of the caudal lobes. Slides from cranial and caudal lobes (n=20) were double-stained (CK8+18 and Ki67) and triple-stained (CK8+18, Ki67, and CD31). The Ki67 proliferation index was calculated using automated tissue and cell segmentation software and compared with manual counting by two independent observers. A substantial variation was seen in the number of Ki67-positive hepatocytes in the different specimens in both double and triple staining (range, 0-50). Correlation coefficients between manual counting and the digital analysis were 0.76 for observer 1 (p <0.001) and 0.78 for observer 2 (p <0.001) with double staining and R(2) = 0.91 for observer 1 and R(2) = 0.89 for observer 2, p <0.001 with triple staining. In conclusion, in rabbit, the hepatocellular proliferation index can be reliably determined using automated tissue and cell segmentation software in combination with IHC multiple staining. Our findings may be useful in clinical practice when Ki67 proliferation index yields prognostic significance
AB - Determination of hepatocyte proliferation activity is hampered by the presence of Ki67-positive non-parenchymal cells. We validated a multicolor immunohistochemical (IHC) approach using multispectral tissue and cell segmentation software. Portal vein branches to the cranial liver lobes of 10 rabbits were embolized, leading to atrophy of the cranial lobes and hyperplasia of the caudal lobes. Slides from cranial and caudal lobes (n=20) were double-stained (CK8+18 and Ki67) and triple-stained (CK8+18, Ki67, and CD31). The Ki67 proliferation index was calculated using automated tissue and cell segmentation software and compared with manual counting by two independent observers. A substantial variation was seen in the number of Ki67-positive hepatocytes in the different specimens in both double and triple staining (range, 0-50). Correlation coefficients between manual counting and the digital analysis were 0.76 for observer 1 (p <0.001) and 0.78 for observer 2 (p <0.001) with double staining and R(2) = 0.91 for observer 1 and R(2) = 0.89 for observer 2, p <0.001 with triple staining. In conclusion, in rabbit, the hepatocellular proliferation index can be reliably determined using automated tissue and cell segmentation software in combination with IHC multiple staining. Our findings may be useful in clinical practice when Ki67 proliferation index yields prognostic significance
KW - Animals
KW - Cell Compartmentation
KW - Cell Proliferation
KW - Hepatocytes/cytology
KW - Humans
KW - Immunohistochemistry
KW - Keratin-18/metabolism
KW - Keratin-8/metabolism
KW - Ki-67 Antigen/metabolism
KW - Liver/cytology
KW - Platelet Endothelial Cell Adhesion Molecule-1/metabolism
KW - Rabbits
KW - Software
KW - Staining and Labeling
U2 - https://doi.org/10.1369/0022155412461154
DO - https://doi.org/10.1369/0022155412461154
M3 - Article
C2 - 22941418
SN - 0022-1554
VL - 61
SP - 11
EP - 18
JO - Journal of Histochemistry and Cytochemistry
JF - Journal of Histochemistry and Cytochemistry
IS - 1
ER -