Biberthaler P, Athelogou M, Langer S, Luchting B, Leiderer R, Messmer K
Institute for Surgical Research, Klinikum der Ludwig-Maximilians-Universität Grosshadern, Munich, Germany.
Eur J Med Res. 2003 Jul 31;8(7):275-82.
Transmission electron micrographs are widely used to demonstrate tissue damage. However, the results are qualitative and dependent on the experience of the investigator. Recently, a new multiscale object-based quantitative image analyzing systems (Cellenger) has been introduced to study highly textured black-and-white images. It is unknown, whether this system permits the quantitative image analysis of electron micrographs of parenchymal tissue. Therefore, we analyzed whether the Cellenge system permits the quantitative evaluation of electron micrographs of murine liver under normal conditions and after ischemia-reperfusion injury. The results were compared with those obtained by conventional qualitative classification. -
Transmission electron micrographs from murine liver that had been exposed to isolated reversible ischemia at hypothermic conditions of 4 degrees C, 15 degrees C, 26 degrees C and 37 degrees C, and of sham-operated animals, which served as controls (2 images per animal, n = 3 in each group), were analyzed qualitatively by an investigator with experience in electron microscopy. For quantitative analysis, the Cellenger was used and the following damage parameters were studied: ratio of area of endothelial cell nucleus to area of endothelial cell (N/C ratio), ratio of area of hepatocellular vacuoles to area of total hepatocyte cytoplasm (V/C ratio) and ratio of area of microvilli in the space of Disse to area of the sinusoids (M/S ratio). All values were sampled within one group (n=6) and the data given in [%] (MW +/- SEM). P-values were accepted as significant below 0.05.
After normothermic ischemia, all quantitative damage parameter were significantly altered as compared to sham-operated animals (N/C 15 +/- 9% vs. 37 +/- 7%, V/C 18 +/- 4% vs. 0, and M/S 0 vs. 10 +/- 1%) and all hypothermia groups. The qualitative electron micrograph section analysis corresponded very well with these results.
We demonstrate that an multiscale object-based quantitative analysis of transmission electron micrographs from mouse liver under control conditions and after I/R provide accurate classification of relevant tissue damage parameter. The system is now ready to use for further applications within the field of highly textured electron micrographs.
透射电子显微镜图像被广泛用于显示组织损伤。然而，结果是定性的，且取决于研究者的经验。最近，一种新的基于多尺度对象的定量图像分析系统（Cellenger）已被引入用于研究高度纹理化的黑白图像。目前尚不清楚该系统是否允许对实质组织的电子显微镜图像进行定量图像分析。因此，我们分析了Cellenge系统是否允许在正常条件下以及缺血再灌注损伤后对小鼠肝脏的电子显微镜图像进行定量评估。将结果与通过传统定性分类获得的结果进行比较。
对在4℃、15℃、26℃和37℃低温条件下经历孤立性可逆缺血的小鼠肝脏以及作为对照的假手术动物的透射电子显微镜图像（每组n = 3，每只动物2张图像），由一位有电子显微镜经验的研究者进行定性分析。对于定量分析，使用Cellenger并研究以下损伤参数：内皮细胞核面积与内皮细胞面积之比（N/C比）、肝细胞空泡面积与总肝细胞细胞质面积之比（V/C比）以及狄氏间隙微绒毛面积与窦状隙面积之比（M/S比）。所有值均在一组内采样（n = 6），数据以 [%] 给出（均值±标准误）。P值在0.05以下被认为具有显著性。
与假手术动物（N/C 15±9% 对 37±7%，V/C 18±4% 对 0，M/S 0 对 10±1%）以及所有低温组相比，常温缺血后所有定量损伤参数均有显著改变。定性电子显微镜切片分析与这些结果非常吻合。
我们证明，对对照条件下以及缺血再灌注后小鼠肝脏的透射电子显微镜图像进行基于多尺度对象的定量分析，能够准确分类相关组织损伤参数。该系统现已准备好用于高度纹理化电子显微镜图像领域的进一步应用。
