Nanobarcoding: detecting nanoparticles in biological samples using in situ polymerase chain reaction

Area of Science:

• Biomedical Engineering
• Nanotechnology
• Molecular Biology

Background:

• Determining nanoparticle (NP) fate and biodistribution is crucial for nanomedicine.
• Existing NP biodistribution detection methods have limited sensitivity.
• A novel nanobarcoding method using in situ polymerase chain reaction (ISPCR) is proposed for enhanced NP detection.

Purpose of the Study:

• To demonstrate the proof of concept for a nanobarcoding method for improved NP detection.
• To evaluate the feasibility of nanobarcoding for NP biodistribution studies.
• To validate the nanobarcoding technique in an in vitro cellular model.

Main Methods:

• Nanobarcoded superparamagnetic iron oxide nanoparticles (NB-SPIONs) were synthesized and characterized.
• Cellular uptake was assessed using confocal microscopy with fluorescently labeled NB-SPIONs.
• The nanobarcoding method was validated using solution-phase PCR and ISPCR in HeLa cells.

Main Results:

• NB conjugation stabilized SPIONs and covered their surface, confirmed by DLS and hyperspectral imaging.
• Solution-phase and pseudo-ISPCR confirmed exclusive amplicon generation from NB-SPIONs in a dose-dependent manner.
• ISPCR detected NB-SPION signals within HeLa cells across large areas, despite minimal uptake observed via confocal microscopy.

Conclusions:

• The nanobarcoding method shows proof of concept for enhanced NP detection in vitro.
• Further development is required to standardize the nanobarcoding technique for widespread use.
• This method holds potential for improving NP biodistribution assessments in nanomedicine.