Journal: bioRxiv

Article Title: Towards a scaled-up T cell-mediated cytotoxicity assay in 3D cell culture using microscopy

doi: 10.1101/842039

Figure Lengend Snippet: a) MC38 cells grown in 35mm culture dishes were pulsed with the SIINFEKL peptide (left panels) or unpulsed (right panels) before staining with an antibody against SIINFEKL bound to H2Kb (shown in red). Cell nuclei were stained with Hoechst. b) MC38 spheroids in collagen gel were seeded in 3D cell culture chips. The SIINFEKL peptide was added to the gel with the spheroids before seeding the chips (top row), only to the media in the side channels (middle row), or no peptide was added (bottom row). Sample were stained with the anti-SIINFEKL-bound H2Kb antibody (red) and Hoechst (white) and imaged at 2 hours post-peptide addition (left panels) and at 24 hours post-peptide addition (right panels). Imaging was performed on the Keyence BZ-X710 with a 20X objective.

Article Snippet: The stain solution contained a 1:200 dilution of an antibody against H-2Kb bound to SIINFEKL conjugated to PE (#141603, Biolegend, San Diego, CA) to detect SIINFEKL peptide being presented at the cell surface, Hoechst to stain the cell nuclei, diluted in PBS with 20% FBS (Atlas Biologicals).

Techniques: Staining, Cell Culture, Imaging