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Three-Dimensional Microvasculature in Rat and Human Hearts Using a Non-injection Ca2+-ATPase Method on Thick and Ultra-Thick Sections

Published online by Cambridge University Press:  22 April 2014

Amaiak Chilingaryan ,
Amayak M. Chilingaryan ,
Mikhail Chilingaryan  and
Gary G. Martin
Amaiak Chilingaryan*
Affiliation:
Department of Biology, Occidental College, 1600 Campus Road, Los Angeles, CA 90041, USA Department of Neurology, LAC+USC Medical Center, 1100 N State St, Los Angeles, CA 90033, USA
Amayak M. Chilingaryan
Affiliation:
Orbeli Institute of Physiology, Yerevan, 0028, Armenia
Mikhail Chilingaryan
Affiliation:
Orbeli Institute of Physiology, Yerevan, 0028, Armenia 221 S. Glendale Avenue, Glendale, CA 91205, USA
Gary G. Martin
Affiliation:
Department of Biology, Occidental College, 1600 Campus Road, Los Angeles, CA 90041, USA
*
*Corresponding author. chilingamd@yahoo.com
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Abstract

Currently there are no methods available for staining rat and human myocardial microvasculature on thick sections that would allow for specific staining and differentiation of arterioles, venules, and capillaries. A non-injection technique is described that allows for labeling of the microvascular bed (MVB) in formalin-fixed pieces of the myocardium from humans and the white rat Rattus norvegicus, as well as human full-mount pericardium. Vessel staining is based on the activity of phosphatases (ATPases) and the precipitation of the released phosphate with calcium ions at high pH (pH 10.5–11.5). The resulting precipitate subsequently is converted to black or brown lead sulfide. The specificity of this reaction to vessels of the MVB allows arterioles, venules, capillaries, and pre- and postcapillaries to be clearly visualized in thick (60–100 µm) and ultra-thick (300–500 µm) sections against an unstained background of muscle and connective tissue. In addition, smooth muscle cells of arterioles are also stained allowing for differentiation between arteriolar and venular beds. These observations have not been reported in rat or human myocardium using other methods. This procedure should benefit studies of coronary microcirculation in experimental and pathological conditions, as well as in pharmacological investigations.

Keywords

microcirculationheartATPasecalciumcoronarystaining
Type
Biological Applications
Information
Microscopy and Microanalysis , Volume 20 , Issue 3 , June 2014 , pp. 895 - 902
Copyright
© Microscopy Society of America 2014 

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