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

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
© 2000 The Society of Thoracic Surgeons

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

Jeffrey S. Martin, MD^a, John G. Byrne, MD^a, Olivier Y. Ghez, MD^a, Umer Sayeed-Shah, MD^a, Sergey D. Grachev, MD^a, Rita G. Laurence, BS^a, Lawrence H. Cohn, MD^a

^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@bics.bwh.harvard.edu

Background. We developed a prosthetic left ventricle (LV) tocoronary sinus (CS) shunt (LVCSS) that is autoregulating andprovides LV-powered retrograde perfusion of the coronary sinus.

Methods. Each of 20 Yorkshire pigs underwent 1 hour of leftanterior descending diagonal artery occlusion followed by 3hours of reperfusion. The controls (n = 5) did not have shunttreatment. The LVCSS group (n = 9) underwent shunt treatmentduring the ischemic period. The LVCSS with partial coronarysinus occlusion (PCSO) group (LVCSS+PCSO, n = 6) underwent shunttreatment and PCSO during the ischemic period. Vital stainingand planimetry techniques were used to determine the area atrisk for infarction and the area of necrosis.

Results. The area at risk was not significantly different amonggroups. The area of necrosis was decreased by 53% in the LVCSSgroup 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 acutecoronary artery occlusion. The addition of PCSO to LVCSS furtherimproves myocardial salvage.

Tim Morken

Lab Vision / NeoMarkers

www.labvision.com

-----Original Message-----
From: Bernard Ian R SSgt 59 CRES/MSROP [mailto:Ian.Bernard@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