Fig. 6.

Double staining of cells isolated from bone marrow aspirates of four mamma carcinoma patients for claudin 3 and cytokeratin. The upper panels show in red claudin- and in green cytokeratin-specific staining of bone marrow cells of breast carcinoma patients BM1117, BM412, BM520, BM573 with corresponding bright field images. Air-fixed slides were prepared from mononucleated cells and stained with chicken anti-CLDN3.3 peptide IgG, and the mouse anti-cytokeratin antibody A45/B/B3 detected by Cy3-labeled anti-chicken and FITC-labeled anti-mouse antibodies, respectively. The lower left panels show claudin 3- and cytokeratin-specific staining of CAMA-1 breast cancer cells as positive control and the corresponding bright fields. Chicken control IgG, and mouse MOPC21 antibody were used as negative controls. Exposure times with Cy3 filter were 100 ms, with FITC filter 500 ms, and bright field 1 ms.