SUMMARY: Our increasing ability to resolve fine details using light microscopy is matched by an increasing need to quantify images in order to detect and measure phenotypes. Despite their central role in cell biology, many image analysis tools require a financial investment, are released as proprietary software, or are implemented in languages not friendly for beginners, and thus are used as black boxes. To overcome these limitations, we have developed PyJAMAS, an open-source tool for image processing and analysis written in Python. PyJAMAS provides a variety of segmentation tools, including watershed and machine learning-based methods; takes advantage of Jupyter notebooks for the display and reproducibility of data analyses; and can be used through a cross-platform graphical user interface or as part of Python scripts via a comprehensive application programming interface. AVAILABILITY AND IMPLEMENTATION: PyJAMAS is open-source and available at https://bitbucket.org/rfg_lab/pyjamas. SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online.

Statistical analysis of genetic changes within cell nuclei that are far from the primary tumor would help determine whether such changes have occurred prior to tumor invasion. To determine whether the gene amplification in cells is morphologically and/or genetically related to the primary tumor requires quantitative evaluation of a large number of cell nuclei from continuous meaningful structures such as milk-ducts, tumors, etc., located relatively far from the primary tumor. To address this issue, we have designed an integrated image analysis software system for high-throughput segmentation of nuclei. Filters such as Beltrami flow-based reaction-diffusion, directional diffusion, etc., were used to pre-process the images resulting in a better segmentation. The accurate shape of the segmented nucleus was recovered using an iterative "shrink-wrap" operation. The study of two cases of ductal carcinoma in situ in breast tissue supports the biological observation regarding the existence of a preferential intraductal invasion, and therefore a common origin, between the primary tumor and the gene amplification in the cell-nuclei lining the ductal structures in the breast.

Spatial events largely determine the biology of cells, tissues, and organs. In this paper, we present a tool for the quantitative spatial analysis of heterogeneous cell populations, and we show experimental validation of this tool using both artificial and real (mammary gland tissue) data, in two and three dimensions. We present the refined relative neighborhood graph as a means to establish neighborhood between cells in an image while modeling the topology of the tissue. Then, we introduce the M function as a method to quantitatively evaluate the existence of spatial patterns within one cell population or the relationship between the spatial distributions of multiple cell populations. Finally, we show a number of examples that demonstrate the feasibility of our approach.