As Knuth said,
We should forget about small efficiencies, say about 97% of the time: premature optimization is the root of all evil.
IMHO, 90% of your optimization should occur at design stage, based on percieved current, and more importantly, future requirements. If you have to take out a profiler because your application doesn't scale to the required load you have left it too late, and IMO will waste a lot of time and effort while failing to correct the problem.
Typically the only optimizations that are worthwhile are those that gain you an order of magnitude performance improvement in terms of speed, or a multiplier in terms of storage or bandwidth. These types of optimizations typically relate to algorithm selection and storage strategy, and are extremely difficult to reverse into existing code. They may go as deep as influencing the decision on the language in which you implement your system.
So my advice, optimize early, based on your requirements, not your code, and look to the possible extended life of your app.