Forgive my ignorance - I'm a lowly orthopaedic surgery resident in need of some basic science advice! I'm currently involved with a project where we need to measure the longitudinal growth of the proximal tibia in immature rabbits.
We're using a novel technique to stimulate the growth plate and we hypothesize that the rate of endochondral growth in the experimental animals will be greater than that of the controls. We're not interested in calculating a rate; we just need to prove that at the end of our experiment (six weeks) the experimental group outgrew the controls.
We have decided to use fluorochromes to aid our measurement. The plan is to administer alizarin complexone at time 0 and then calcein at time 6 weeks.
Here are my questions:
1. Are those choices of fluorochromes appropriate? - we had originally considered oxytetracycline...can anyone comment on one choice over another?
2. My most important question - after six weeks can I expect to see two distinct bands on longitudinal sectioning or will the band corresponding to time 0 already have undergone resorption?
3. I have no personal experience preparing the fluorochromes for injection...can anyone comment on what it takes to get it from the box it was packaged in to a form appropriate for injecting into an animal? I've read a few protocols that make it seem like it's difficult to handle - buffering it, storing/preparing it under cold conditions, filtering it, etc.