Journal:

Article Title: Retinal pigment epithelial cells phagocytosis of T lymphocytes: possible implication in the immune privilege of the eye

doi:

Figure Lengend Snippet: Photomicrograph of Hoechst staining of 24 hour co-culture hRPE cells/T lymphocytes. RPE cells (200 000 cells/well) were allow to adhere overnight at 37°C in 24 well flat bottomed tissue culture plates in complete medium, before allogeneic purified T lymphocytes (500 000 T cells/well) were added. After 24 hours of co-culture, Hoechst 33342, a bisbenzimidazole DNA binding dye was added to the culture at 10 μg/ml, for 30 minutes at 37°C. The supernatant was discarded, and cells trypsinised to disrupt cell-matrix and cell-cell binding, dislodging lymphocytes just attached and not ingested by RPE cells. Thereafter, the collected cells were washed, cytospun, and visualised under ultraviolet light. On the photomicrograph RPE cells (large dark blue nucleus and large cytoplasm) have phagocytosed T cells (small light blue nucleus and no cytoplasm). Magnification ×400.

Article Snippet: Hoechst staining After 24 hours of co-culture, Hoechst 33342, a bisbenzimidazole DNA binding dye (Boehringer Mannheim GmbH) was added to the culture at 10 μg/ml, for 30 minutes at 37°C.

Techniques: Staining, Co-Culture Assay, Purification, Binding Assay