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. 1998 Nov;80(5):509–516. doi: 10.1136/hrt.80.5.509

Intravascular stents: a new technique for tissue processing for histology, immunohistochemistry, and transmission electron microscopy

N Malik 1, J Gunn 1, C Holt 1, L Shepherd 1, S Francis 1, C Newman 1, D Crossman 1, D Cumberland 1
PMCID: PMC1728851  PMID: 9930055

Abstract

Background—Study of the vascular response to stent implantation has been hampered by difficulties in sectioning metal and tissue without distortion of the tissue stent interface. The metal is often removed before histochemical processing, causing a loss of arterial architecture. Histological and immunohistochemical sections should be 5 µm with an intact tissue stent interface.
Objectives—To identify the most suitable cutting and grinding equipment, embedding resin, and slides for producing thin sections of stented arteries with the stent wires in situ for histological, immunohistochemical, and transmission electron microscopic (TEM) analyses.
Methods—20 balloon stainless steel stents were implanted in the coronary arteries of 10 pigs. Twenty eight days later the stented arterial segments were excised, formalin fixed, embedded in five different resins (Epon 812, LR white, T9100, T8100, and JB4), and sectioned with two different high speed saws and a grinder for histological, immunohistochemical, and TEM analyses. Five stented human arteries were obtained at necropsy and processed using the best of the reported methods.
Results—The Isomet precision saw and grinder/polisher unit reliably produced 5 µm sections with most embedding resins; minimum section thickness with the horizontal saw was 400 µm. Resin T8100, a glycol methacrylate, enabled satisfactory sectioning, grinding, and histological (toluidine blue, haematoxylin and eosin, and trichromatic and polychromatic stains) and immunohistochemical analyses (α smooth muscle actin, von Willebrand factor, vimentin, proliferating cell nuclear antigen, and CD68 (mac 387)). T9100 and T8100 embedded stented sections were suitable for ultrastructural examination with TEM. Stented human arterial sections showed preserved arterial architecture with the struts in situ.
Conclusion—This study identified the optimal methods for embedding, sawing, grinding, and slide mounting of stented arteries to achieve 5 µm sections with an intact tissue metal interface, excellent surface qualities, histological and immunohistochemical staining properties, and suitability for TEM examination. The technique is applicable to experimental and clinical specimens.

 Keywords: stents;  resin;  histology;  immunohistochemistry;  transmission electron microscopy

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Figure 1  .

Figure 1  

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Histological staining of stented arterial sections. (A) Haematoxylin and eosin stained section (scale bar = 500 µm) showing preserved arterial wall architecture and stent struts in situ; and (B) at higher magnification (scale bar = 100 µm) with an intact internal elastic lamina (arrowheads). (C) Toluidine blue stained section (scale bar = 100 µm) showing varying spatial orientation of cells within the neointimal layer. (D) Trichromatic stained section (scale bar = 100 µm) showing struts in situ, preserved arterial architecture, and an intact internal elastic lamina (arrowheads); and (E) at higher magnification (scale bar = 50 µm) showing cells around the strut arranged in clusters, unlike cells nearer the lumen. (F) Haematoxylin stained section (scale bar = 5 µm) showing an intact tissue metal interface with neointimal cells around the strut arranged in clusters.

Figure 2  .

Figure 2  

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Immunohistochemical staining of stented arterial sections. (A) α Smooth muscle actin staining (scale bar = 100 µm): α smooth muscle actin positive cells are stained red (New Fuchsin Red), the counterstain is methyl green. All cells within the neointima, with distinctly differing spatial orientation, and media are positive for a smooth muscle actin. (B) von Willebrand Factor staining (scale bar = 50 µm): endothelium (arrowheads) of the vasa vasorum of the adventitia is stained red (New Fuchsin Red), the counterstain is methyl green.

Figure 3  .

Figure 3  

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Transmission electron micrographs of stented coronary artery sections. (A) Neointimal layer 28 days after stenting with abundant extracellular matrix and interspersed smooth muscle cells (scale bar = 0.5 µm) arranged in distinctly different spatial orientation. Arrowhead indicates the luminal surface. (B) Phenotype of the neointimal cells is confirmed as smooth muscle cells (scale bar = 0.18 µm). Note characteristic cytoplasmic motor regions of these cells (arrowhead).

Figure 4  .

Figure 4  

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Stented human arterial sections. (A) Stented left anterior descending artery (LAD) section (scale bar = 500 µm), without histological staining showing preserved arterial wall architecture with an eccentric calcified plaque and stent struts in situ. (B) Haematoxylin and eosin stained section (scale bar = 50 µm) showing the stent strut embedded into relatively normal part of the arterial wall (arrowhead). (C) Haematoxylin and eosin staining showing complex calcified plaque (p) with an intact capsule (scale bar = 50 µm). Note position of the guidewire (arrowhead) in the arterial lumen. (D) Unstained LAD section, obtained at necropsy from another patient, showing preserved arterial architecture and stent struts in situ. The dissection (arrowhead) mostly involves the relatively normal area of the arterial wall. Note the adjacent calcified plaque (p).

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These references are in PubMed. This may not be the complete list of references from this article.

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