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Cell Painting, a high-content image-based assay for morphological profiling using
multiplexed fluorescent dyes
#MMPMID27560178
Bray MA
; Singh S
; Han H
; Davis CT
; Borgeson B
; Hartland C
; Kost-Alimova M
; Gustafsdottir SM
; Gibson CC
; Carpenter AE
Nat Protoc
2016[Sep]; 11
(9
): 1757-74
PMID27560178
show ga
In morphological profiling, quantitative data are extracted from microscopy
images of cells to identify biologically relevant similarities and differences
among samples based on these profiles. This protocol describes the design and
execution of experiments using Cell Painting, which is a morphological profiling
assay that multiplexes six fluorescent dyes, imaged in five channels, to reveal
eight broadly relevant cellular components or organelles. Cells are plated in
multiwell plates, perturbed with the treatments to be tested, stained, fixed, and
imaged on a high-throughput microscope. Next, an automated image analysis
software identifies individual cells and measures ?1,500 morphological features
(various measures of size, shape, texture, intensity, and so on) to produce a
rich profile that is suitable for the detection of subtle phenotypes. Profiles of
cell populations treated with different experimental perturbations can be
compared to suit many goals, such as identifying the phenotypic impact of
chemical or genetic perturbations, grouping compounds and/or genes into
functional pathways, and identifying signatures of disease. Cell culture and
image acquisition takes 2 weeks; feature extraction and data analysis take an
additional 1-2 weeks.
free
free
free
