Using Cell-ID 1.4 with R for Microscope-Based Cytometry

ALAN BUSH; ARIEL CHERNOMORETZ; RICHARD YU; ANDREW GORDON; ALEJANDRO COLMAN LERNER

Current protocols in molecular biology

Wiley-Interscience

Lugar: New York; Año: 2011

1934-3639

This unit describes a method for quantifying various cellular features (e.g., volume, total and subcellular fluorescence localization) from sets of microscope images of individual cells. It includes procedures for tracking cells over time. One purposefully defocused transmission image (sometimes referred to as bright-field or BF) is acquired to segment the image and locate each cell. Fluorescent images (one for each of the color channels to be analyzed) are then acquired by conventional wide-field epifluorescence or confocal microscopy. This method uses the image processing capabilities of Cell-ID and data analysis by the statistical programming framework R (R-Development-Team, 2008), which we have supplemented with a package design to analyze Cell-ID output. Both programs are open-source software packages.