Jonker A, Geerts W J, Charles R, Lamers W H, Van Noorden C J
Department of Anatomy and Embryology, University of Amsterdam, Academic Medical Center, The Netherlands.
J Histochem Cytochem. 1995 Oct;43(10):1027-34. doi: 10.1177/43.10.7560880.
To analyze regional differences in the activity of glutamate dehydrogenase in rat liver in situ, we developed an image recording and processing system for monitoring the formation of a colored final reaction product in time. All absorbance measurements of test and control reactions in time in consecutive sections were used to fit the data to a quadratic curve, with the derivative at t = 0 representing the initial velocity of formazan formation. The images of sections incubated for test and control reactions were topographically matched with an affine transformation using the positions of vessels as fiducials. Specific enzyme activity was calculated by subtracting the coefficients representing the initial velocity at corresponding locations in the test and control reactions and appeared to be 8 and 4 mumoles glutamate converted per min per cm3 of tissue at 20 degrees C in pericentral and periportal zones of fasted female rats, respectively. Those values are in agreement with biochemical data. The ability to construct two-dimensional images of cellular distribution patterns of enzyme activity in liver lobules is particularly useful for the study of metabolic zonation in this organ.
为了分析大鼠肝脏原位谷氨酸脱氢酶活性的区域差异,我们开发了一种图像记录和处理系统,用于实时监测有色终产物的形成。对连续切片中测试反应和对照反应随时间的所有吸光度测量数据进行二次曲线拟合,t = 0 时的导数代表甲臜形成的初始速度。使用血管位置作为基准,通过仿射变换将测试反应和对照反应孵育后的切片图像进行地形匹配。通过减去代表测试反应和对照反应相应位置初始速度的系数来计算比酶活性,在禁食雌性大鼠的中央静脉周围区和门静脉周围区,20℃时每立方厘米组织每分钟分别有8和4微摩尔谷氨酸转化。这些值与生化数据一致。构建肝小叶酶活性细胞分布模式二维图像的能力对于研究该器官的代谢分区特别有用。