Fluorescence Microscopy Imaging Calibration for Quantifying Nanocarrier Binding to Cells During Shear Flow Exposure

Area of Science:

• Biomedical Engineering
• Nanotechnology
• Pharmacology

Background:

• Targeted drug delivery utilizes nanocarriers for drug transport, a rapidly advancing healthcare field.
• Understanding nanocarrier binding dynamics is crucial for optimizing nanocarrier design, especially for smaller particles.
• Current microscopy techniques face limitations in quantifying binding dynamics for sub-micron nanocarriers due to pixel size constraints.

Purpose of the Study:

• To develop and validate a novel calibration technique for quantitative measurement of nanocarrier binding dynamics.
• To enable precise analysis of nanocarrier-cell interactions, particularly for nanoparticles smaller than microscopy pixel resolution.
• To assess the temporal binding profile of antibody-targeted nanocarriers under fluid shear stress.

Main Methods:

• A novel calibration technique combining radioisotope counting and fluorescence imaging was developed.
• The technique was applied to quantify the binding dynamics of antibody-targeted nanocarriers to endothelial cells.
• Nanoparticle binding was assessed under conditions of fluid shear stress.

Main Results:

• The novel calibration technique enabled quantitative determination of nanocarrier binding dynamics.
• Accurate measurements were achieved for nanoparticles smaller than a microscopy image pixel.
• The temporal profile of endothelial cell binding for two antibody-targeted nanocarrier types was characterized.

Conclusions:

• The developed radioisotope counting and fluorescence imaging calibration technique overcomes limitations of traditional microscopy for nanocarrier analysis.
• This method provides a robust platform for quantitative assessment of nanocarrier binding dynamics, crucial for optimizing targeted drug delivery systems.
• The findings facilitate the design and development of more effective nanocarrier-based therapeutics.