A Versatile Cell Death Screening Assay Using Dye-Stained Cells and Multivariate Image Analysis

A Versatile Cell Death Screening Assay Using Dye-Stained Cells and Multivariate Image Analysis.

A novel dye-based method for measuring cell death in image-based screens is presented. Unlike conventional high- and medium-throughput cell death assays that measure only one form of cell death accurately, using multivariate analysis of micrographs of cells stained with the inexpensive mix, red dye nonyl acridine orange, and a nuclear stain, it was possible to quantify cell death induced by a v...

mRNA analysis of intracytoplasmically-stained, FACS-purified pancreatic islet cells using the quantitative nuclease protection assay

A high-throughput, multiplex cell death assay using an RNAi screening approach.

This protocol outlines a high-throughput, multiplex cell death assay and its use in conjunction with a genome-scale siRNA screen to identify genes that cooperate with a drug to induce apoptosis. The assay, ApoLive-Glo (Promega), measures viability of drug-treated, reverse-transfected cells via the fluorescent CellTiter-Fluor reagent, which includes a substrate that is cleaved by a live cell pro...

Automatic Quantification of Immunohistochemically Stained Cell Nuclei Using Unsupervised Image Analysis

A method for quantification of images of immunohistochemically stained cell nuclei by computing area proportions is presented. The image is transformed by a principal component transform. The resulting first component image is used to segment the objects from the background using dynamic thresholding of the P2/A-histogram, where P2/A is a global roundness measure. Then the image is transformed ...

A versatile multivariate image analysis pipeline reveals features of Xenopus extract spindles

Imaging datasets are rich in quantitative information. However, few cell biologists possess the tools necessary to analyze them. Here, we present a large dataset of Xenopusextract spindle images together with an analysis pipeline designed to assess spindle morphology across a range of experimental conditions. Our analysis of different spindle types illustrates how kinetochore microtubules ampli...