[Histonet] Help; looking for a stain receipe for TTC

[Bernard Ian R SSgt 59 CRES/MSROP]

Hello folk or fellow histonetters(in particular in research), 
 
I'm in the process of replicating the histochemical staining method in a
protocol article titled " LV-Powered Coronary Sinus Retroperfusion Reduces
infarct Size in Acutely Ischemic Pigs."
 
The made reagents for staining the heart tissue calls for a pre staining is
a Phthalol-blue, which we will be substituting with Evans Blue.  I need a
concentration and receipe to make this up?   The article does not provide
this.  The article states that the " ascending aorta was occluded and
phthalo-blue(Evans Blue) was injected into the left ventricle.
 
Finally the article states: " The left Ventricle was sliced, parallel to the
atriopventricular groove, into several 5-10 mm slices and incubated for 30
mins at 37 deg C in TTC or Triphenyltetrazolium Chloride.  There are 2
concentrations by Sigma, which one is to be used here, a 95% or 99.5 %?
Need receipe to make this solution?
 
What is Trisma HCL and Trisma Base?  I need catalog #'s and source?
 
The article goes on to state for results:That "the Mycardial tissue
containing dehydrogenase enzymes is stained brick red by TTC, but areas of
necrotic tissue depleted of these enzymes are not stained and appear as
white or pale yellow."
 
 
Thanks
SSgt Bernie
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