Hi guys! For my current master's project, I am designing a high throughput screening of different small molecules to enhance the function of Batf3-dependent dendritic cells. The readouts include the ability of these DCs to cross-present OVA peptides/proteins to CD8+ T cells. We plan to do the screening on a 384 well-plate platform with the maximal volume of 80uL per well. But I am wondering:
1. What are the numbers of DCs and T cells should I put into each well?
2. What kind of plate should I suse? V or U or flat bottom?
3. At the staining step, should I wash the stain or not. If so, how?
4. Is there any literature concerning this issue that I could read?
Thanks all!
Hi Minh..this is surprising for me that you have designed all these conditions without any literature support or performing any trial experiments beforehand. I have worked with DC-T cells experiments long back, and have worked with 98-well plates. My experiments were different, but giving some suggestions.
- I would suggest U-bottom plate, as there are better chances are cells interacting with each other and growth of cells.
-Attaching some literature, probably, you can get some clue from there.
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