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High Throughput Fluorometric Technique for Assessment of Macrophage Phagocytosis and Actin Polymerization

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Quantifying fluorescent nanoparticle uptake in mammalian cells using a plate reader.

Hye Ji Shin¹, Minjeong Kwak², Sihwa Joo³

¹Biometrology Group, Division of Chemical and Biological Metrology, Korea Research Institute of Standards and Science, 267 Gajeong-Ro, Yuseong-Gu, Daejeon, 34113, Republic of Korea.

Scientific Reports

|November 23, 2022

Summary

Researchers developed a simple, quantitative assay to measure nanoparticle uptake in cells using a plate reader. This method addresses common issues, offering a robust tool for nanoparticle research.

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Area of Science:

Nanotechnology
Biotechnology
Cell Biology

Background:

Nanoparticle applications are expanding rapidly, necessitating tools to study their biological interactions.
Existing methods for measuring nanoparticle cellular uptake are often labor-intensive and lack quantitative accuracy.

Purpose of the Study:

To develop an accessible, robust, and quantitative assay for measuring nanoparticle cellular uptake.
To optimize the assay to minimize variations and interferences for reliable results.

Main Methods:

Developed a quantitative assay using a plate reader for fluorescently labeled nanoparticles.
Normalized fluorescence intensity to optical density to account for cell number variations.
Optimized washing steps and sample handling temperature to reduce residual nanoparticle interference and nanoparticle efflux.

Main Results:

Successfully demonstrated the assay with fluorescently labeled 50 nm polystyrene beads in Jurkat cells.
Confirmed the assay's applicability in A549 (lung carcinoma) and RPMI8226 (lymphocyte) cell lines.

Conclusions:

The developed plate reader assay provides an easily accessible and robust method for quantifying nanoparticle cellular uptake.
This assay offers a valuable tool for researchers investigating nanoparticle-biological entity interactions across various cell types.