The short answer is yes, to be completely sure it will look the same you need to test with all browsers.
The longer answer is that Firefox, Chrome and Opera all have good standards compliance so if it works in one, it's very likely to work in the others.
IE is a minefield of its own and the differences between IE6, 7 and 8 can be pronounced. I usually try and avoid supporting IE6 altogether now so don't bother to check it and IE8 has an IE7 backward compatibility mode so you can test both 7 and 8 together. If you need IE6, Microsoft provide Virtual Machine images with it installed on XP so you can test against it rather than trying to hack multiple versions of IE onto your machine.
As a final check something like browsershots.org can help you see what it will look like on a wide variety of OS/browser combos.
One thing I found very useful is using a CSS framework like Yahoo's YUI — they have taken the hit of getting stuff working consistently across lots of browsers so you can leverage their work with a bit of additional styling of your own.