Quantifying the level of nanoparticle uptake in mammalian cells using flow cytometry

Area of Science:

• Biomedical Engineering
• Cell Biology
• Nanotechnology

Background:

• Accurate nanoparticle uptake quantification is crucial for medicine and environmental science.
• Conventional methods like electron microscopy and confocal imaging are time-consuming and semi-quantitative.

Purpose of the Study:

• To develop and validate flow cytometry-based assays for quantifying fluorescent nanoparticle uptake in mammalian cells.
• To establish a quantitative, routine-applicable method for nanoparticle uptake analysis.

Main Methods:

• Developed two flow cytometry assays: percentage of nanoparticle-containing cells and Molecules of Equivalent Soluble Fluorophore (MESF) using calibration beads.
• Applied assays to quantify fluorescent silica nanoparticle uptake in A549 lung carcinoma cells.
• Tested assays across nine different mammalian cell types to assess cell type/size correlations.

Main Results:

• Successfully quantified nanoparticle uptake in A549 cells, moving beyond semi-quantitative measures.
• Demonstrated the utility of flow cytometry for routine, quantitative nanoparticle uptake analysis.
• Investigated and identified correlations between cell characteristics and nanoparticle uptake efficiency.

Conclusions:

• Flow cytometry provides a robust and quantitative method for measuring nanoparticle uptake in mammalian cells.
• The developed assays are versatile and applicable to various cell types, facilitating broader research.
• These assays enable more accurate assessment of nanoparticle interactions with cells for diverse applications.