Home › Cell Biology › Fluorescent Nanoparticles for the Measurement of Ion Concentration in Biological Systems
Cell Biology JoVE (Open Access) Citable · DOI

Fluorescent Nanoparticles for the Measurement of Ion Concentration in Biological Systems

DOI: 10.3791/2896-v
What you'll learn
  • ✓ Synthesize fluorescent sodium nanosensors for ion concentration imaging
  • ✓ Inject nanosensors intracellularly and subcutaneously in biological systems
  • ✓ Measure and optimize nanosensor fluorescent response to ion concentrations
  • ✓ Acquire in vivo and intracellular sodium imaging data
Protocol

Fluorescent nanoparticles produced in our lab are used for imaging ion concentrations and ion fluxes in biological systems such as cells during signaling and interstitial fluid during physiological homeostasis.

Difficulty
advanced
Total time
~3–5 days (synthesis, characterization, and in vivo imaging per animal)
Model organism
Not explicitly specified; likely rodent based on subcutaneous/intracellular injection context
Biosafety
BSL-1

Steps

1
Prepare optode solution for nanosensor synthesis

Formulate the optode solution components required as the base reagent for fluorescent nanoparticle production.

▶ 01:26
2
Create sodium-selective nanosensors

Synthesize fluorescent nanoparticles using the optode solution to produce sodium nanosensors with ion-selective properties.

▶ 01:55
3
Determine optimal intracellular nanosensor response

Characterize nanosensor fluorescence output across varying sodium concentrations to establish calibration and selectivity parameters.

▶ 03:15
4
Perform intracellular nanosensor injection

Introduce nanosensors directly into cells using microinjection or similar technique to enable intracellular ion imaging.

▶ 04:59
5
Perform subcutaneous nanosensor injection

Inject nanosensors into interstitial fluid via subcutaneous administration to measure ion concentrations in physiological compartments.

▶ 05:40
6
Acquire in vivo and intracellular sodium images

Collect fluorescence microscopy images of nanosensors from both intracellular and in vivo subcutaneous sites to visualize sodium concentrations.

▶ 07:03

🚨 Failure Case Library (1) + Submit your own case

moderate
Abnormal fluorescence localization (aggregates / wrong compartment)
Fluorescence forms aggregates inside cells, or localizes to a compartment different from the expected native pattern.
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