van der Steen A F, Thijssen J M, van der Laak J A, Ebben G P, de Wilde P C
Biophysics Laboratory of the Institute of Ophthalmology, Nijmegen, The Netherlands.
J Microsc. 1994 Jul;175(Pt 1):21-33. doi: 10.1111/j.1365-2818.1994.tb04784.x.
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 micrometers and 10 micrometers 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 x 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 micrometers. 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 x 250 micrometers. 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.
一种强大的新方法被用于研究生物组织的光学显微镜特性与声学特性之间的相关性。将肝脏标本切成连续的切片，厚度分别为250微米和10微米。对厚切片进行声学研究，对薄切片进行光学显微镜观察。使用可通过光学和声学检测及定位的标记物，在声学和光学切片（2.5×2.5毫米）中找到并重建相应区域。从声学研究的切片中重建参数图像。声学参数包括30兆赫兹时的衰减、衰减谱斜率（10至50兆赫兹之间）、30兆赫兹时的背向散射、背向散射谱斜率（10至50兆赫兹之间）以及局部超声速度。在10至50兆赫兹的频率范围内获取声学图像，横向分辨率约为50微米。用于光学显微镜观察的切片按照戈德纳三色染色技术进行染色。使用数字图像分割技术定量确定组织学组成。计算250×250微米区域内富含胶原蛋白的纤维组织、管腔结构和间隙空间的百分比以及细胞核数量。将这些组织学特征与相邻切片中相应区域获得的声学参数相关联。由此有可能找出对声学参数负责的组织学成分。
