I need to arrest the heart in diastole prior to fixation to analyze cardiomyocyte size using WGA. I have seen publications using 40 mM KCl, but I have concerns that such a hypotonic solution will burst the cells. Anyone have a protocol they want to share? I was thinking of make a 40 mM KCl solution in PBS and using 30 mM BDM. Any thoughts? Thanks in advance.
St Thomas' Cardioplegic solution No.2 (David Hearse's development) has a 16 mM K+ conc and is delivered at 4 - 8 degrees C. We've generally used this for cardiac arrest and preservation during transplant. The histological appearnce of the heart tissur is of a publishable high quality. You can also make up this this high K+ solution as a modified Krebs' buffer.
Other papers have reported using a higher K+ conc of 25 mM at 22 degrees C.
In rats, "With a median thoracotomy, the heart was vented via the right atrium, and 30 mL of a 1.5% solution of KCl in PBS was infused through the carotid artery to arrest the heart". Please see my publication: Failure of Calcineurin Inhibitors to Prevent Pressure-Overload Left Ventricular Hypertrophy in Rats. Circulation Research1999;84:722-728 doi: 10.1161/01.RES.84.6.722
It is downloadable at http://circres.ahajournals.org/content/84/6/722.full
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