Home › Cell Biology › Localization and Relative Quantification of Carbon Nanotubes in Cells with Multispectral Imaging Flow Cytometry
Cell Biology JoVE (Open Access) Citable · DOI

Localization and Relative Quantification of Carbon Nanotubes in Cells with Multispectral Imaging Flow Cytometry

DOI: 10.3791/50566-v
What you'll learn
  • ✓ Prepare water-dispersible carbon nanotubes for cellular uptake studies
  • ✓ Acquire multispectral imaging flow cytometry data using ImageStream
  • ✓ Analyze CNT localization and quantification using IDEAS software
Protocol

In this study, the main purpose was to monitor the cellular uptake and eventual exocytosis of carbon nanotubes (CNTs). From this perspective, we proposed here a unique method using multispectral imaging flow cytometry allowing a quantification and localization of CNTs in a statistically relevant number of cells.

Difficulty
advanced
Total time
~4–6 hours (sample preparation through acquisition and analysis)
Biosafety
BSL-1

Steps

1
Prepare water dispersible carbon nanotubes

Disperse carbon nanotubes in aqueous solution using appropriate surfactants or chemical modification to enable cellular uptake studies. Ensure homogeneous suspension before cell exposure.

▶ 01:38
2
Acquire cells with ImageStream multispectral imaging

Load prepared cell samples into the ImageStream flow cytometer and acquire multispectral image data to capture CNT localization and cellular morphology.

▶ 03:24
3
Analyze CNT localization using IDEAS software

Process acquired ImageStream data with IDEAS analysis software to quantify CNT internalization, determine subcellular localization, and generate statistical metrics across the cell population.

▶ 04:07
4
Interpret CNT uptake and exocytosis results

Review representative analyses of CNT internalization patterns to assess cellular uptake efficiency and track exocytosis dynamics over experimental timepoints.

▶ 08:52
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