I recently submitted my application to New York without the barcode separators. I know some countries do have the requirement but I could not find any details whether it was needed for US applications or not.
From what I’ve heard on this forum the presence of barcode separators is not going to affect the application, just make sure your pack is organized logically and easy to navigate.
To be honest the separators add more confusion because they are generic so they have education and TB sections which aren’t necessarily a requirement on this visa for most US citizens. Plus there is an accommodation separator and this it the sponsor documents separator talks about accommodation - confusing if you ask me
For this reason I created a contents letter and organized the pack in that order. But I really don’t even think that is a requirement.
But I haven’t got my visa yet so I wouldn’t base your decision on me, but if you don’t use them at least I’m in your boat too