A new device for the efficient pulverisation and extraction of myocardial biopsies for high energy phosphate analysis.
We developed a new device for processing frozen myocardial biopsies. Frozen samples of 20 to 50 mg were dropped into a 25 ml stainless steel centrifuge tube held in a custom-made aluminium container precooled in liquid nitrogen. A stainless steel pestle attached to a stainless steel disk was driven by a modified heavy-duty staple gun to pulverise the tissue rapidly at low temperatures. The tissue powder was extracted with 0.3N PCA at 0 degree C in the centrifuge tube which was then transferred to a Sorvall super-speed centrifuge. Values for adenosine triphosphate (ATP) were 5.6 +/- 0.7 mumol . g-1 wet weight (mean +/- SD). Creatine phosphate (CP) yield was 12.2 +/- 3 mumol . g-1 wet weight. The % recovery of an added internal standard for ATP was 86 +/- 18% and for CP 90 +/- 16% with the new method.
D000255:Adenosine Triphosphate / Q000032:analysis*; D000818:Animals; D002309:Cardiology / Q000295:instrumentation; D004285:Dogs; D005615:Freezing; D008722:Methods; D009206:Myocardium / Q000032:analysis*; D010725:Phosphocreatine / Q000032:analysis*
D010725:Phosphocreatine; D000255:Adenosine Triphosphate