CD8T细胞四聚体染色效率分析
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CD8 staining eYciency. DTH biopsy from patient 5 (lesion DC + 3 peptides) stained with HLA-A2.1-tyrosinase 369 tetramer (red), anti-CD8 (green) and DAPI (blue) with increasing exposure time for the FITC Wlter (ranging from 2–3 s (a) to 15–20 s (c), for comparison: exposure time TRITC 7–8 s). Arrows indicate CD8+ cells that were not visible at shorter exposure times for the FITC Wlter. In c, only for the boxed area an increased exposure time (15–20 s) is shown to prevent overexposure from other parts of the section. d Enlarged two color overlay for green (CD8) and blue (DAPI) of the boxed area from c. Tetramer analysis by Xow cytometry of DTH-derived T cells from patient 5. On the y-axis MHC class I tetramer PE staining and on the x-axis CD4 (e) or CD8 (f). Note the diVerence in Xuorescence intensity in the tetramer positive population (MFI CD4:152 vs. MFI CD8:83) whereas the percentage of positive cells is equal (17%). CD8+ cells visualized with the ABC-peroxidase kit with AEC (red-brown staining) as substrate in corresponding areas on tissue sections from tonsil Wxed with acetone (g) or PFA (h)
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