Quantitation of oncogene products by computer-assisted image analysis and flow cytometry.
The use of antibodies permits the study of oncogene product expression in cells and tissues. However, quantitation of the levels of expression in immunohistochemical preparations is beset by difficulties, and the available scoring system provide semiquantitative data at best. Here we describe the use of computer-assisted image analysis for determination of oncoprotein levels in a model system and compare the results with those generated by flow cytometric analysis. The oncogene products measured are located in the nucleus (c-myc p62 and c-fos p55), the inner surface of the membrane (c-ras p21), and both sides of the membrane (c-erbB-2 p185). In each instance, both analytic modalities yielded concordant results. Our data indicate that computer-assisted image analysis is a useful tool for quantitating cell components in immunohistochemical preparations.
Czerniak
B
B
Department of Pathology, Montefiore Medical Center, Albert Einstein College of Medicine, Bronx, New York 10467.
Herz
F
F
Wersto
R P
RP
Alster
P
P
Puszkin
E
E
Schwarz
E
E
Koss
L G
LG
D001943:Breast Neoplasms / Q000032:analysis* / Q000378:metabolism / Q000473:pathology; D002460:Cell Line; D005434:Flow Cytometry; D015870:Gene Expression; D006801:Humans; D007091:Image Processing, Computer-Assisted; D007150:Immunohistochemistry; D015513:Oncogene Proteins / Q000032:analysis* / Q000378:metabolism; D011518:Proto-Oncogene Proteins / Q000235:genetics / Q000378:metabolism; D016760:Proto-Oncogene Proteins c-fos; D016271:Proto-Oncogene Proteins c-myc; D016283:Proto-Oncogene Proteins p21(ras); D018719:Erb-b2 Receptor Tyrosine Kinases; D014407:Tumor Cells, Cultured / Q000032:analysis / Q000378:metabolism / Q000473:pathology
D015513:Oncogene Proteins; D011518:Proto-Oncogene Proteins; D016760:Proto-Oncogene Proteins c-fos; D016271:Proto-Oncogene Proteins c-myc; D016283:Proto-Oncogene Proteins p21(ras); D018719:Erb-b2 Receptor Tyrosine Kinases; C501009:HRAS protein, human