Figure 3. Intact photoreceptors in culture initially dissociated using three different techniques. After 2 days in culture, cells were immunostained with anti-rhodopsin (A, E, I; green) for outer segment labeling and with DAPI nuclear staining (B, F, J; blue) and observed under epifluorescence illumination. A-D: After 2 days in culture, cells dissociated using the enzymatic treatment with gentle pipeting (Protocol A) still showed the outer segment attached to the nucleus/cell body; however, it developed a circular shape. E-H: In cells dissociated using the enzymatic treatment with gentle pipeting followed by centrifugation (Protocol B) rhodopsin was mostly expressed within the cytoplasm of the cells. I-L: When dissociating the cells with the non-enzymatic treatment with gentle pipeting (Protocol C) rhodopsin expression was observed in outer segment debris. Expression of rhodopsin and DAPI within a same cell was rare. The scale bars represent 5 μm.