A new method to correlate acoustic spectroscopic microscopy (30 MHz) and light microscopy

Abstract

Summary A powerful new method is used to investigate the correlation between light microscopic and acoustic properties of biological tissues. Specimens of liver were sectioned into successive slices, 250 μm and 10 μm thick. The thick sections were investigated acoustically, the thin sections by means of light microscopy. Markers that could be detected and located, both optically and acoustically, were used to find and reconstruct corresponding regions in the acoustic and optical sections (2·5 × 2·5 mm). Parameter images were reconstructed from the sections investigated acoustically. The acoustic parameters were attenuation at 30 MHz, the slope of the attenuation spectrum (between 10 and 50 MHz), backscattering at 30 MHz, the slope of the backscattering spectrum (between 10 and 50 MHz) and the local ultrasound velocity. Acoustic images were obtained in the frequency range from 10 to 50 MHz, yielding a lateral resolution of about 50 μm. The sections for light microscopy were stained according to the Goldner trichrome staining technique. The histological composition was determined quantitatively, using digital image segmentation techniques. The percentage of collagen‐rich fibrous tissue, luminal structure and interstitial spaces, and the number of nuclei were calculated for regions of 250 × 250 μm. These histological features were correlated with the acoustic parameters obtained from the corresponding regions in adjacent sections. It was thus possible to find the histological components responsible for acoustic parameters.

Correlation of histology and acoustic parameters of liver tissue on a microscopic scale

“…In this study, the correlation between local spectral acoustic parameters and local histological features in liver was investigated by a method developed by van der Steen et al (1994). This method correlates the acoustic properties of a 250 µm thick section of biologica} tissue to the light microscopie features of an adjacent 10 µm thick section.…”

“…The nuclear density had no influence on any acoustic parameter. Some caution should be exercised with this conclusion, because it may be possible that the nuclear density varies significantly within 250 µm in the tissue, in which case the employed method is not able to produce a consistent result (van der Steen et al 1994).…”

“…The methods (van der Steen et al 1994) describe a technique to correlate the acoustic properties of a tissue section to the light microscopie features in an adjacent section. The acoustic part of the methods was inspired by the work of F. S. Poster (Poster et al 1984;D' Astous and Poster 1986).…”

“…Recently, a method has been developed to identify the histological features responsible for acoustic tissue characterization parameters (van der Steen et al 1994).…”