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.