I am trying to couple angiogenesis and osteogenesis in a rodent model of spinal cord injury. What I would like to do is take a bone which exhibits osteoporosis (bone loss) and couple this with the absence of type H vessels by staining for CD31 and Endomucin.
Is it possible to fix a bone in formalin, then microCT scan it the next day, and then decalcify the same bone and perform IHC / histology on it without compromising results?
Also, are there any better stains you would recommend to analyse bone vasculature?
At least you can try once or twice to prove on. It is difficult task but not impossible
CT does not interfere with IHC. Take into account that it is better to decalcify with EDTA than with strong acids, in order to preserve proteins. CD31 is OK, you could add CD34 or ERG but, in this context, I think CD31 is OK.
Pablo
Any damage from x-rays is going to happen on the molecular or atomic scale. In living organisms this can definitely pose a problem (i.e. a strand break in a DNA molecule), but you're not going to see any physical changes in fixed tissue. Repeated exposure to radiation can eventually degrade materials but that takes a lot of dose and is not a concern from a standard ex vivo microCT.
Perhaps consider perfusing with a CT contrast agent (I'm thinking a tungsten-based one to try and get contrast above cortical bone). You may get a good-enough visualization of the vasculature and you don't have to worry about co-registration.
Joseph R Merrill Pablo Roitman Luis Rodrigo thank you all very much for your help, it is most appreciated.
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