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Updated: Jul 30, 2026
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Image-based Flow Cytometry Technique to Evaluate Changes in Granulocyte Function In Vitro
Published on: December 26, 2014
Quantitation of AgNORs by flow versus image cytometry
1Equipe RFMQ, Laboratoire TIMC UMR CNRS 5525, Institut Albert Bonniot, Université Joseph Fourier, La Tronche, France. bjacquet@imag.fr
Summary
This study introduces flow cytometry (FCM) for quantifying silver-stained nucleolar organizer regions (AgNORs), offering a faster, reproducible alternative to image analysis. This method accurately assesses cell proliferation and cycling speed in tumors.
Area of Science:
- Cell biology
- Biotechnology
- Cancer research
Background:
- Nucleolar organizer regions (AgNORs) are nucleolar proteins that bind silver salts.
- AgNOR size, measured by image analysis (ICM), correlates with cell cycle time and cancer prognosis.
- ICM is time-consuming and lacks reproducibility across different laboratories.
Purpose of the Study:
- To develop a novel flow cytometry (FCM) method for AgNOR quantitation.
- To establish a rapid and reproducible technique for assessing cell proliferation and cycling speed.
Main Methods:
- Utilized FCM to measure the decrease in forward scattered light caused by silver precipitates.
- Correlated FCM measurements with AgNOR relative area determined by ICM.
- Performed double staining with Ki-67/MIB 1 antigen and silver nitrate for FCM analysis.
Main Results:
- Silver precipitates significantly decreased forward scattered light in FCM.
- FCM-based AgNOR quantitation showed a strong correlation with ICM measurements.
- Successfully analyzed cell proliferation markers (Ki-67/MIB 1) and AgNORs simultaneously using FCM.
Conclusions:
- FCM provides a valid, rapid, and reproducible method for AgNOR quantitation.
- This technique enables accurate comparison of growth fraction and cycling speed in proliferating cell populations, including tumors.
