Does Twitter ratio of followers vs. following really matter?

No, it doesn't matter!

What really matters is that you're happy with the number of people you're following, that you're not overwhelmed by the number of tweets you get. If you feel you are following some people you don't really know who do a huge number of tweets that you don't really find interesting you might want to think about unfollowing some of them, but I think if either you find their tweets interesting or they don't tweet too often you might as well keep them. Sure, it makes you look more popular if you have more followers than you are following, but who wants to get hung up on that? At the moment I've got almost twice as many followers as I'm following (100 vs 53), but that's because I don't automatically follow everyone who follows me, and some of the people I follow tweet quite a lot, and that's the amount of tweets I want coming in. If those people didn't tweet so much I'd follow some more, and I wouldn't worry about my ratio. A low ratio could just mean you follow a lot of popular, interesting tweeters who don't follow back even 1% of their huge number of followers, rather than a lot of people who'll follow anyone back because they don't have many followers and want to keep hold of any they get.