Improved quantitative characterization of atherosclerotic plaque composition with immunohistochemistry, confocal fluorescence microscopy, and computer-assisted image analysis

Abstract

To quantitatively characterize contributions of major constituents to the composition of a given atherosclerotic plaque, we have developed an approach employing immunohistochemistry, confocal scanning laser microscopy, and computer-assisted image analysis. The method developed permits identification of plaques that are particularly vulnerable to rupture and elucidation of the nature of the composition of a given plaque, as well as the extent of luminal encroachment. Thus, it should be useful in experimental animals and ultimately in patients in delineating compositional changes in response to potentially deleterious genetic and environmentally induced factors and to potentially therapeutic interventions designed to diminish plaque vulnerability.

Raman Spectroscopic Evaluation of the Effects of Diet and Lipid-Lowering Therapy on Atherosclerotic Plaque Development in Mice

“…Novel techniques to improve the quantitative characterization of plaque composition are under development. 20 However, such histological or immunohistochemical evaluations still require sectioning, and quantification of atherosclerosis remains difficult: Computer-assisted quantification of positively stained structures in histological sections can be obscured by the nonspecific staining of adventitial fat and by overlying tissue that hinders staining of structures in deeper layers. Moreover, decalcification before sectioning and staining leads to inaccurate visualization and quantification of CS that are present in the plaque.…”

Attenuation of Neointimal Vascular Smooth Muscle Cellularity in Atheroma by Plasminogen Activator Inhibitor Type 1 (PAI-1)

“…The captured digital images were transferred to a Dell 400 NT workstation for cropping and processing to yield 24-bit RGB images per section isolated on a white background as previously described (Taatjes et al 2000). The processed images were opened in MetaMorph Image Analysis software for analysis.…”

Hematopoietic origin of hepatic stellate cells in the adult liver

“…To analyze the presence of lipid droplets in EGFP ϩ cells, we performed ORO staining combined with immunofluorescence assays. 27,28 Subpanels i and ii in Figure 3C show the light microscopic image and the fluorescent microscopic image of ORO ϩ cells, respectively. The majority of these ORO ϩ cells were positive for EGFP (Figure 3Ciii).…”

“…Visualization of lipid with ORO staining was obtained by excitation with the 647-nm laser line. 28 Images were acquired using FV10-ASW software (Ver 1.3; Olympus) and processed using Adobe Photoshop CS. …”