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Related Experiment Video

Updated: Jun 1, 2026

Workflow for High-content, Individual Cell Quantification of Fluorescent Markers from Universal Microscope Data, Supported by Open Source Software
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Published on: December 16, 2014

Computer-based fluorescence quantification: a novel approach to study nucleolar biology.

Mohamed Kodiha1, Piotr Bański, Ursula Stochaj

  • 1Department of Physiology, McGill University, 3655 Promenade Sir William Osler, Montreal, H3G 1Y6, Canada.

BMC Cell Biology
|June 7, 2011
PubMed
Summary

We developed new quantitative microscopy methods to precisely measure nucleolar protein and RNA dynamics. These techniques enable high-throughput screening for a deeper understanding of nucleolar functions in cellular processes.

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Last Updated: Jun 1, 2026

Workflow for High-content, Individual Cell Quantification of Fluorescent Markers from Universal Microscope Data, Supported by Open Source Software
09:57

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Published on: December 16, 2014

Using Computer Vision Libraries to Streamline Nuclei Quantification
06:25

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Published on: June 6, 2025

Quantitative Immunofluorescence to Measure Global Localized Translation
09:13

Quantitative Immunofluorescence to Measure Global Localized Translation

Published on: August 22, 2017

Area of Science:

  • Cell Biology
  • Molecular Biology
  • Biophysics

Background:

  • Nucleoli are essential nuclear compartments involved in ribosome biogenesis and regulation of key cellular processes like cell cycle and apoptosis.
  • The nucleolus's dynamic proteome composition and macromolecular flux present challenges in linking molecular changes to cellular outcomes.
  • Understanding nucleolar dynamics is crucial for deciphering its role in health and disease, including tumorigenesis and stress responses.

Purpose of the Study:

  • To develop and validate quantitative immunofluorescence methods for analyzing nucleolar dynamics.
  • To enable the study of nucleolar protein and RNA kinetics under various physiological conditions.
  • To facilitate high-throughput screening of nucleolar components and activities.

Main Methods:

  • Quantitative immunofluorescence combined with computer-based image analysis.
  • Monitoring dynamic association of proteins and RNA with nucleoli.
  • Measuring de novo RNA synthesis within nucleoli.

Main Results:

  • Demonstrated effectiveness in monitoring dynamic protein and RNA association with nucleoli.
  • Quantified stress-dependent changes in nucleolar protein concentrations (endogenous and GFP-tagged).
  • Successfully measured nucleolus-associated de novo RNA synthesis.

Conclusions:

  • The developed protocols allow quantitative analysis of nucleolar protein kinetics at single-cell and high-throughput levels.
  • Compatibility with automated high-throughput screening (HTS) platforms enables large-scale data acquisition.
  • These advancements in quantitative microscopy are expected to yield new insights into complex nucleolar functions.