[Histonet] Whole mount porcine trachea staining

[Griffin, Michelle A]

Would anyone have an idea about how to stain whole porcine trachea so
that the cartilage rings show prominently? There have been several
excellent articles that use Alcian Blue or an Alcian Blue/Alizarin Red S
combination to stain rodent tracheas (mouse/rat) and the pictures are
wonderful (Schipani et al., 2001; Regnier et al., 2002; Dolle et al.,
1993). However, I have tried to reproduce the results using the same
protocols just a different species and am having no luck (I did try out
each method with some mouse tracheas in my lab and the results mimicked
what I saw in published articles). I have tried different
digestion/clearing solutions, times, pH, etc and continue to end up with
a trachea that is quite ugly with no differentiation between cartilage
rings and connective tissue. 

Does anyone have a suggestion on why pig trachea would not stain even
similar to mouse trachea? Any suggestions would be greatly appreciated.
Thanks so much for your time and input. 

Michelle A. Griffin BA, BS, MHA
University of Iowa
Comparative Pathology Laboratory
200 Hawkins Drive
143 Medical Research Center
Iowa City, IA 52242
319-384-4620