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Analyzing Cellular Internalization of Nanoparticles and Bacteria by Multi-spectral Imaging Flow Cytometry

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Published on: June 8, 2012

Quantifying Nanoparticle Internalization Using a High Throughput Internalization Assay.

Sarah K Mann^1,2,3, Ewa Czuba^1,3, Laura I Selby^1,3

¹Drug Delivery, Disposition and Dynamics, Monash Institute of Pharmaceutical Sciences, Monash University, Parkville, Victoria, 3052, Australia.

Pharmaceutical Research

|July 6, 2016

Summary

This summary is machine-generated.

A new DNA sensor, the Specific Hybridization Internalization Probe (SHIP), tracks nanoparticle uptake in cells. SHIP reveals faster nanoparticle internalization in 3T3 cells than CEM cells via a dynamin-dependent pathway.

Keywords:

endocytosis inhibitor internalization nanoparticles sensor

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Published on: September 28, 2022

Area of Science:

Biotechnology
Cell Biology
Nanomedicine

Background:

Nanoparticle internalization is key for drug delivery.
Understanding nanoparticle uptake mechanisms is crucial for developing effective nanotherapeutics.

Purpose of the Study:

To develop a DNA molecular sensor, the Specific Hybridization Internalization Probe (SHIP), to investigate nanoparticle internalization kinetics and mechanisms.
To differentiate between cell-surface associated and internalized nanoparticles.

Main Methods:

Utilized self-assembling polymeric 'pHlexi' nanoparticles functionalized with a Fluorescent Internalization Probe (FIP).
Studied interactions with 3T3 and CEM cell lines.
Employed chemical inhibitors (sodium azide, Dyngo-4a, EIPA) to elucidate internalization pathways.

Main Results:

Nanoparticle internalization kinetics were significantly faster in 3T3 cells (90% internalized) compared to CEM cells (20% internalized).
Nanoparticle uptake was identified as a dynamin-dependent process.
Internalized nanoparticles were trafficked to lysosomal compartments.

Conclusions:

The SHIP assay effectively distinguishes cell-surface bound from internalized nanoparticles.
SHIP can probe nanoparticle internalization kinetics and mechanisms, aiding in the design of improved nanoparticle delivery systems.
SHIP is a simple, high-throughput technique with broad applications in therapeutic delivery research.