Tumor cell counting using an image analysis program for MIB-1 immunohistochemistry.
A cell counting method for MIB-1 immunohistochemistry, using an image analysis program, NIH Image, on a personal computer, has advantages over manual cell counting by microscopy. MIB-1 slides were photographed at a magnification of x 50 on 24 x 36 mm color films and the photographs were then enlarged to adequate size for observation. The MIB-1-positive cells, on the enlargements, were marked with a white pen, and negative cells with a black pen. Subsequently, the image of an enlargement was converted into data that can be processed by the program using a flatbed image scanner. Threshold levels were adjusted according to the respective white and black markings, and the program automatically counted the number of MIB-1-positive and negative tumor cells. Our method has the following significant advantages: Once cells had been marked and the image converted into the NIH Image data, the cell count process was completed within a few minutes; in contrast to manual microscopic cell counting, count overlapping or cells missed by the observer are eliminated; overlapping nuclei could be separated by the marking procedure; our method required no elaborate and costly image analysis system. The present method may be recommended to researchers undertaking cell kinetic studies utilizing immunohistochemical methods.
Ide
M
M
Department of Neurosurgery, Tokyo Women's Medical College Dai-ni Hospital.
Jimbo
M
M
Yamamoto
M
M
Kubo
O
O
D001932:Brain Neoplasms / Q000378:metabolism* / Q000473:pathology; D002452:Cell Count / Q000379:methods*; D005909:Glioblastoma / Q000378:metabolism* / Q000473:pathology; D006801:Humans; D007091:Image Processing, Computer-Assisted; D007150:Immunohistochemistry