High Resolution Quantitative Imaging

Project title: High Resolution Quantitative Imaging of Tissue Pathology

Project summary: Histopathology is limited in its utility at the point of care. Fluorescence microscopy can be used to acquire real-time images of tissue morphology and with appropriate algorithms can rapidly quantify features associated with disease. For example, in situ tissue imaging using fluorescent stains may be useful for intraoperative detection of residual cancer in surgical tumor margins. However, robust methods for segmentation and quantitative analysis of heterogeneous images are essential to enable automated diagnosis. Thus, the goal of this project is to obtain high resolution imaging of tissue morphology through employing fluorescence microscopy and vital fluorescent stains and to develop a quantitative strategy to segment and quantify tissue features in heterogeneous images, such as nuclei and the surrounding stroma, which will enable automated diagnosis of thick tissues. Toward this end, we have developed automated algorithms that enable us to segment nuclei and nucleoli, which we collectively refer to as fluorescent positive features (FPFs) and quantify their size and density. This information can be used to quantitatively diagnose tissue, which can be useful for a variety of applications. In particular, we have demonstrated the ability of this combination of techniques to diagnose excised human breast tissue. Representative images are shown in the figure below.

 High Resolution Quantitative Imaging. Representative  confocal fluorescence images of adipose tissue, ductal  carcinoma in situ (DCIS), and invasive ductal carcinoma (IDC)  are shown in A through C, respectively. D-F: FPFs segmented  using an image processing algorithm called maximally stable  extremal regions (MSER) are false colored green and overlaid  onto the raw confocal fluorescence image. G-I: Histologic  slides with H&E staining show similar histology to confocal  images in A-C. Slides were prepared with the same specimens  from which confocal images were acquired. Scale bar is 100  µm.