Quantitative characterization of preneoplastic progression using single‐cell computed tomography and three‐dimensional karyometry

Abstract

The development of morphological biosignatures to precisely characterize preneoplastic progression necessitates high-resolution three-dimensional (3D) cell imagery and robust image processing algorithms. We report on the quantitative characterization of nuclear structure alterations associated with preneoplastic progression in human esophageal epithelial cells using single-cell optical tomography and fully automated 3D karyometry. We stained cultured cells with hematoxylin and generated 3D images of individual cells by mathematically reconstructing 500 projection images acquired using optical absorption tomographic imaging. For 3D karyometry, we developed novel, fully automated algorithms to robustly segment the cellular, nuclear, and subnuclear components in the acquired cell images, and computed 41 quantitative morphological descriptors from these segmented volumes. In addition, we developed algorithms to quantify the spatial distribution and texture of the nuclear DNA. We applied our methods to normal, metaplastic, and dysplastic human esophageal epithelial cell lines, analyzing 100 cells per line. The 3D karyometric descriptors elucidated quantitative differences in morphology and enabled robust discrimination between cell lines on the basis of extracted morphological features. The morphometric hallmarks of cancer progression such as increased nuclear size, elevated nuclear content, and anomalous chromatin texture and distribution correlated with this preneoplastic progression model, pointing to the clinical use of our method for early cancer detection. ' 2010 International Society for Advancement of CytometryKey terms preneoplastic progression; Barrett's esophagus; esophageal cancer; single cell; computed tomography; 3D karyometry; image processing NUCLEAR architecture is a key factor in cell functioning and pathogenesis (1). The functional relevance of nuclear structure in many core cell processes necessitates detailed studies on the interplay between nuclear architecture and functions like gene expression. One of the most important applications of three-dimensional (3D) karyometric studies is the quantitative characterization of the morphological changes associated with malignancy. Alterations in nuclear structure hold high clinical and research relevance, especially in the context of early cancer detection (2,3). Although much remains to be learned about the molecular mechanisms that drive cancerrelated alterations in nuclear structure and the higher-order spatial organization of chromatin, accurate quantification of structural parameters could be beneficial for developing robust biosignatures for early cancer detection.Rapid advances in automation sciences and imaging technologies have facilitated the use of computer-aided karyometry, where sets of morphological descriptors (referred to as morphometric signatures) are extracted from microscopy images using digital image processing algorithms. Morphometric signatures generated from a statistically significant number of cells may then be used to p...

Inverse Scattering Shape Reconstruction of 3d Bacteria Using the Level Set Algorithm

“…For lower values of SNR, obtaining meaningful reconstruction results of any target using almost any method will be challenging. In the future, the level set algorithm will be examined using experimental data in the high frequency, once made available from systems similar to those reported in [5,6].…”

“…Advances in micro-fabrication and micro-fluidics had facilitated the development of systems to allow 3D tomography of a single cell [5,6]. In the cell tomography system developed by Meyera et al, cell samples were rotated from 0 • to 360 • in steps of 0.72 • in order to achieve 500 projections [5].…”

“…In the cell tomography system developed by Meyera et al, cell samples were rotated from 0 • to 360 • in steps of 0.72 • in order to achieve 500 projections [5]. A three dimensional filtered back-projection algorithm was implemented to reconstruct the shape of cancerous cells [6].…”

“…As reported in [12], the BWA are most effectively delivered as aerosols in air and they represent a serious threat because they are invisible and tasteless. However, investigating bacteria immersed in other media is also important for validation with controlled laboratory measurements similar to those conducted in [5,6]. The later will be the subject of future work.…”

“…Fig. 1 shows a synthetic system to collect the scattering fields at different incident and receiving directions (angles) and at multiple high frequencies, conceptually similar to the single cell system in [5,6]. The single cell system reported in [5] demonstrated that the 3D shape reconstruction increased the accuracy of cancer cells detection three fold, compared with the 2D shape reconstruction.…”