Rychtáriková Renata, Náhlík Tomáš, Shi Kevin, Malakhova Daria, Macháček Petr, Smaha Rebecca, Urban Jan, Štys Dalibor
Institute of Complex Systems, Faculty of Fisheries and Protection of Waters, University of South Bohemia, Zámek 136, 373 33 Nové Hrady, Czech Republic.
Institute of Complex Systems, Faculty of Fisheries and Protection of Waters, University of South Bohemia, Zámek 136, 373 33 Nové Hrady, Czech Republic.
Ultramicroscopy. 2017 Aug;179:1-14. doi: 10.1016/j.ultramic.2017.03.018. Epub 2017 Mar 18.
Current biological and medical research is aimed at obtaining a detailed spatiotemporal map of a live cell's interior to describe and predict cell's physiological state. We present here an algorithm for complete 3-D modelling of cellular structures from a z-stack of images obtained using label-free wide-field bright-field light-transmitted microscopy. The method visualizes 3-D objects with a volume equivalent to the area of a camera pixel multiplied by the z-height. The computation is based on finding pixels of unchanged intensities between two consecutive images of an object spread function. These pixels represent strongly light-diffracting, light-absorbing, or light-emitting objects. To accomplish this, variables derived from Rényi entropy are used to suppress camera noise. Using this algorithm, the detection limit of objects is only limited by the technical specifications of the microscope setup-we achieve the detection of objects of the size of one camera pixel. This method allows us to obtain 3-D reconstructions of cells from bright-field microscopy images that are comparable in quality to those from electron microscopy images.
当前的生物学和医学研究旨在获取活细胞内部详细的时空图谱，以描述和预测细胞的生理状态。我们在此展示一种算法，可根据使用无标记宽场明场光透射显微镜获得的z轴堆叠图像，对细胞结构进行完整的三维建模。该方法可视化的三维物体体积相当于相机像素面积乘以z轴高度。计算基于找到物体扩散函数的两个连续图像之间强度不变的像素。这些像素代表强光衍射、光吸收或发光物体。为实现这一点，使用从雷尼熵导出的变量来抑制相机噪声。使用此算法，物体的检测极限仅受显微镜设置的技术规格限制——我们实现了对单个相机像素大小物体的检测。这种方法使我们能够从明场显微镜图像中获得细胞的三维重建，其质量与电子显微镜图像相当。
