[Quantitative non-radioactive in situ hybridization for measurement of specific mRNA using image analyzer]

To understand cells, information on the state of gene expression in each cell is essential. We determined the copy number of c-myc mRNA in HL60 cell by first carrying out in situ hybridization using thymine-thymine dimerized anti-sense oligonucleotides probes to c-myc mRNA and the hybridized thymine dimers were localized by the immunoperoxidase method. The density of each cell was measured by an image analyzer and converted to the mRNA copy number in each cell, using dot blot hybridization results, as a reference standard. It was found that the numbers of c-myc mRNA copies varied 0-4,000 when expressed at the unit of cell.