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

Tue Oct 14 10:35:22 CDT 2003

SSgt Bernie

If this (below) is the same article, I notice that they have an email given
for contact. Maybe ask them the particulars. 

I can tell you that Trizma HCl and Trizma Base are from Sigma (look under
Trizma in the chemical list). They supply it like this, in two parts, so you
can mix the exact pH buffer you wish to use (between pH 7 and 9) . They give
instructions with the chemicals. Another way to do it is to get Trizma
pre-set crystals which will give you the exact pH you want with out any
mixing of chemicals (between pH 7 and 9).

Ann Thorac Surg 2000;69:84-89
(c) 2000 The <http://ats.ctsnetjournals.org/misc/terms.shtml>  Society of
Thoracic Surgeons 

  _____  

LV-powered coronary sinus retroperfusion reduces infarct size in acutely
ischemic pigs 

Jeffrey S. Martin, MDa, John G. Byrne, MDa, Olivier Y. Ghez, MDa, Umer
Sayeed-Shah, MDa, Sergey D. Grachev, MDa, Rita G. Laurence, BSa, Lawrence H.
Cohn, MDa 
a Division of Cardiac Surgery, Brigham and Women's Hospital, Harvard Medical
School, Boston, Massachusetts, USA 
Address reprint requests to Dr Byrne, Division of Cardiac Surgery, Brigham
and Women's Hospital, 75 Francis St, Boston, MA 02115
e-mail: jgbyrne <@t> bics.bwh.harvard.edu <mailto:jgbyrne <@t> bics.bwh.harvard.edu> 
Background. We developed a prosthetic left ventricle (LV) to coronary sinus
(CS) shunt (LVCSS) that is autoregulating and provides LV-powered retrograde
perfusion of the coronary sinus. 
Methods. Each of 20 Yorkshire pigs underwent 1 hour of left anterior
descending diagonal artery occlusion followed by 3 hours of reperfusion. The
controls (n = 5) did not have shunt treatment. The LVCSS group (n = 9)
underwent shunt treatment during the ischemic period. The LVCSS with partial
coronary sinus occlusion (PCSO) group (LVCSS+PCSO, n = 6) underwent shunt
treatment and PCSO during the ischemic period. Vital staining and planimetry
techniques were used to determine the area at risk for infarction and the
area of necrosis. 
Results. The area at risk was not significantly different among groups. The
area of necrosis was decreased by 53% in the LVCSS group and by 73% in the
LVCSS+PCSO group when compared to controls (p < 0.01 among all groups). 
Conclusions. The LVCSS reduces infarct size in pigs after acute coronary
artery occlusion. The addition of PCSO to LVCSS further improves myocardial
salvage. 

Tim Morken
Lab Vision / NeoMarkers
www.labvision.com

-----Original Message-----
From: Bernard Ian R SSgt 59 CRES/MSROP [mailto:Ian.Bernard <@t> LACKLAND.AF.MIL] 
Sent: Tuesday, October 14, 2003 7:02 AM
To: Histonet (E-mail)
Subject: [Histonet] Help; looking for a stain receipe for TTC
Importance: High

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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