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Figure 2 | Journal of Ovarian Research

Figure 2

From: A high M1/M2 ratio of tumor-associated macrophages is associated with extended survival in ovarian cancer patients

Figure 2

Flow cytometric validation of immunofluorescence analysis of total, M1, and M2 TAMs in ovarian cancer. (A) Cancer islet and stromal regions were microdissected from tissue sections. The obtained tissues were dissociated into cell suspensions and subjected to flow cytometry analysis. The arrows indicate the microdissected cancer tissue. (B) Representative flow cytometry analyses of the cell suspensions obtained from the cancer islet and stromal regions. The cells were stained with PE-conjugated anti-keratin and FITC-conjugated anti-CD68 monoclonal antibodies. The asterisk indicates the percentage of CD68+ TAMs (keratin-) in the total cells, as determined through flow cytometry. (C) Representative flow cytometry analyses of M1 and M2 TAMs in a cell suspension. The cells were stained with PE-conjugated anti-HLA-DR (or anti-CD163, anti-iNOS, and anti-VEGF) and FITC-conjugated anti-CD68 monoclonal antibodies. The asterisk indicates the percentage of M1 or M2 TAMs in the total cell population, as determined through flow cytometry. (D) The correlations between the total, M1, and M2 TAM densities, as determined through immunofluorescence and the TAM ratios in the total cells analyzed through flow cytometry. R2 indicates the square of the Pearson’s product–moment correlation coefficient.

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