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Metal ions can be separated from one another by complexation with organic ligands–the chelating agent– to form uncharged chelates. Here, the chelating agent must contain hydrophobic groups and behave as a weak acid, losing a proton to bind with the metal. Since most organic ligands used in this process are insoluble or undergo oxidation in the aqueous phase, the chelating agent is initially added to the organic phase and extracted into the aqueous phase. The metal-ligand complex is formed in...

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Related Experiment Video

Updated: Jun 24, 2026

Rapid Analysis and Exploration of Fluorescence Microscopy Images

Rapid Analysis and Exploration of Fluorescence Microscopy Images

Published on: March 19, 2014

Extracting rich information from images.

Anne E Carpenter¹

¹Imaging Platform, Broad Institute of Harvard and MIT, 7 Cambridge Center, Cambridge, MA, 02142, USA. anne@broad.mit.edu

Methods in Molecular Biology (Clifton, N.J.)

|April 7, 2009

Summary

This summary is machine-generated.

High-content screening (HCS) generates vast cellular image data. This protocol uses CellProfiler software to extract hundreds of features per cell, unlocking richer biological insights from images.

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Area of Science:

Cell biology
Biotechnology
Bioinformatics

Background:

Automated microscopy enables high-throughput screening, generating massive cellular image datasets.
Traditional analysis often reduces complex image data to single numerical outputs, ignoring rich information.
High-content screening (HCS) acknowledges the high information content within cell images.

Purpose of the Study:

To provide a detailed protocol for analyzing cellular images using open-source CellProfiler software.
To demonstrate the extraction of hundreds of cellular features beyond traditional metrics.
To showcase the application of this method using a cytoplasm-to-nucleus translocation assay example.

Main Methods:

Utilized CellProfiler, an open-source cell image analysis software.
Developed a protocol to measure hundreds of features for individual cells.
Included cell compartment size/shape, staining intensity, and texture analysis.
Applied the protocol to publicly available images from a translocation assay.

Main Results:

Successfully extracted hundreds of quantitative features from individual cells.
Demonstrated detailed analysis of cellular compartments, organelles, and staining characteristics.
Validated the protocol's effectiveness on a relevant biological assay.

Conclusions:

CellProfiler enables comprehensive analysis of high-content screening image data.
This protocol significantly enhances the information extracted from cellular images.
The approach offers a powerful tool for detailed biological investigation in high-throughput experiments.