First, cells are labeled with Anti-ErbB-2 MicroBeads and positively selected on a first separation column. Then ErbB-2
+ cells are fixed and applied to a second MACS® Column, and subjected to intracellular staining of cytokeratin filaments using two alkaline phosphatase-conjugated cytokeratin-specific antibody clones, CK3-11D5 and CK3-3E4. After elution from the column, cells are transferred onto a microscope slide, incubated with the substrate, and can be analyzed by light microscopy. During the entire staining procedure, the cells remain within the magnetic field of the column, thus minimizing cell loss.