Detecting microcalcifications in atherosclerotic plaques by a simple trichromic staining method for epoxy embedded carotid endarterectomies

  • M. Relucenti | michela.relucenti@uniroma1.it Department of Human Anatomy, Sapienza University of Rome, Italy.
  • R. Heyn Department of Human Anatomy, Sapienza University of Rome, Italy.
  • L. Petruzziello Department of Human anatomy, Researcher, Italy.
  • G. Pugliese Department of clinical sciences, II Faculty of Medicine, Sapienza university of Rome, Italy.
  • M. Taurino Department of Vascular Surgery, II Faculty of Medicine, Sapienza University of Rome, Italy.
  • G. Familiari Department of Human Anatomy, Sapienza University of Rome, Italy.

Abstract

Atherosclerotic plaques have a high probability of undergoing rapid progression to stenosis, becoming responsible of acute coronary syndrome or stroke. Microcalcifications may act as enhancers of atherosclerotic plaque vulnerability. Considering that calcifications with a diameter smalller than 10 mm in paraffin embedded tissue are rather difficult to detect, our aim was to analyze microcalcifications on semithin sections from epoxy resin embedded samples of carotid endarterectomies using an original trichromic stain (methylene blue- azur B - basic fuchsine - alizarin red). We have compared samples stained either with our method, methylene blue-azur B alone or with Von Kossa staining, and methylene blue-azur B -basic fuchsine alone or with Von Kossa staining. Our method resulted to be simple and fast (ca. 2 min), it gives a sharp general contrast for all structures and allows to easy identify collagen and elastin. In addition, gray-green colour associated to intracellular lipid droplets evidences foam cells, which are particularly abundant in endarterectomies samples. Mast cells and their metachromatic granules are also well recognized. Calcifications over 0,5 mm are clearly recognizable. In conclusion, microcalcifications are clearly distinguished from the extracellular matrix in spite of their reduced dimensions. Methylene blue-azur B-basic fuchsine-alizarin red method is easy to use, reproducible, and is particularly suitable for the identification of microcalcifications in the morphological analysis of atherosclerotic plaques.

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Author Biographies

M. Relucenti, Department of Human Anatomy, Sapienza University of Rome
Depatrment of Human anatomy, Researcher
R. Heyn, Department of Human Anatomy, Sapienza University of Rome
Depatrment of Human anatomy, Researcher
L. Petruzziello, Department of Human anatomy, Researcher
Depatrment of Human anatomy, PhD Student
G. Pugliese, Department of clinical sciences, II Faculty of Medicine, Sapienza university of Rome

Depatemt of Clinical Sciences

Associate Professor

M. Taurino, Department of Vascular Surgery, II Faculty of Medicine, Sapienza University of Rome

Departmet of Vascular Surgery

Full Professor

G. Familiari, Department of Human Anatomy, Sapienza University of Rome
Department of Human Anatomy, Full Professor
Published
2010-07-14
Section
Technical Notes
Keywords:
Microscopy, Histochemistry, Cytology
Statistics
Abstract views: 703

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How to Cite
Relucenti, M., Heyn, R., Petruzziello, L., Pugliese, G., Taurino, M., & Familiari, G. (2010). Detecting microcalcifications in atherosclerotic plaques by a simple trichromic staining method for epoxy embedded carotid endarterectomies. European Journal of Histochemistry, 54(3), e33. https://doi.org/10.4081/ejh.2010.e33