u/OkGOcategories

Hi everyone! Thanks so much for all of your help :) I have started optimization for a large tetramer screen (18 x MHC-I specific CD8 tetramers) I had initially optimized the screen for 1 tetramer per well, but would like to increase to as many tetramers per well as possible. I had tried to multiplex this to 18 but unfortunately have a lot of what I suspect is nonspecific staining. :( (see images below) I have plans to titrate the PE/tetramer and try at room temp vs. on ice. Is there any other condition that I should try to optimize or does anyone have any tips? I am expecting very few tetramer positive cells in these conditions. Thank you! (also- the cd19 + population looks strange/nonexistant because this protocol requires B cell depletion.) TIA!

https://preview.redd.it/ibtpd5g104uh1.png?width=1980&format=png&auto=webp&s=a41e1f36d13e88b56766939dd389edf22e78d271

Hope everyone is doing well! I am trying to make a steak sandwich (kinda fancy haha) and was wondering where I should tweak/improve. Recipe is posted below. Thanks!

Toasted ciabatta Medium rare ribeye, sliced thinly against the grain Chimichurri Garlic aioli Caramelized shallots Arugula, lightly dressed with lemon + olive oil + salt Shaved Parmesan Pickled red onions (very small amounts) salt + cracked black pepper

A few years ago I took a Sicilian cooking class at Eataly where we made swordfish involtini that was fantastic, and I cannot seem to find the recipe anywhere. It had some pine nuts, basil or parsley I cannot remember, and some golden raisins, among other things. It did not have cheese (of course) or breadcrumbs. If anyone has any leads or a recipe that sounds similar I would deeply appreciate!