Shape features

The purpose of the workflow is ....

First you need

Workflow of data-analysis strategies for image-based cell profiling.

1. Image analysis (image correction, image segmentation, feature extraction)
2. Image quality control
3. Preprocessing extracted features
4. Dimensionality reduction
5. Single-cell data aggregation
6. Measuring profile similarity
7. Assay quality assessment
8. Downstream analysis

Examples of publications where the workflow has been used at least partly:

• Image-based multivariate profiling of drug responses from single cells
• Phenotypic profiling of the human genome by time-lapse microscopy reveals cell division genes
• Mapping genetic interactions in human cancer cells with RNAi and multiparametric phenotyping
• Cell shape and the microenvironment regulate nuclear translocation of NF‐κB in breast epithelial and tumor cells
• Genome-wide RNAi Screening Identifies Protein Modules Required for 40S Subunit Synthesis in Human Cells
• Cell Painting, a high-content image-based assay for morphological profiling using multiplexed fluorescent dyes
• Systematic morphological profiling of human gene and allele function via Cell Painting

The ultimate goal of the NET framework is to make images of networks processable by computers. Therefore we want to have a pixel based image as input, as output we want a representation of the network visible in the image that retains as much information about the original network as possible. NET achives this by first segmenting the image and then vectorizing the network and then extracting information. The information we extract is

• First and foremost the graph of the network. We find the crossings (nodes) and connections between crossings (edges) and therefore extract information about the neighborhood relations, the topology of the network.
• We also extract the coordinates of all nodes which enables us to embed them into space. We therefore extract information about the geometry of the network.
• Last but not least we track the radii of the edges in the extraction process. Therefore every edge has a radius which can be identified with its conductivity.

In the following we will first provide detailed instructions on how to install NET on several platforms. Then we describe the functionality and options of each of the four scripts that make up the NET framework.

The invention comprises a software tool, NeuronMetrics, which functions as a set of modules that run in the open-source program ImageJ. NeuronMetrics features a novel method for estimating neural “branch number” (a measure of the axonal complexity) from two-dimensional images. In addition, the tool features a novel method for modeling neural structure in large “gaps” that result from image artifacts.