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Pharmacology JoVE (Open Access) Citable · DOI

An Integrated Raman Spectroscopy and Mass Spectrometry Platform to Study Single-Cell Drug Uptake, Metabolism, and Effects

DOI: 10.3791/60449-v
What you'll learn
  • ✓ Integrate Raman spectroscopy and mass spectrometry for single-cell drug analysis
  • ✓ Quantify drug uptake and metabolite production at single-cell resolution
  • ✓ Assess cellular metabolic responses to pharmaceutical compounds
Protocol

This protocol presents an integrated Raman spectroscopy-mass spectrometry (MS) platform that is capable of achieving single-cell resolution. Raman spectroscopy can be used to study cellular response to drugs, while MS can be used for targeted and quantitative analysis of drug uptake and metabolism.

Difficulty
advanced
Total time
~3–5 days (cell culture preparation, drug treatment, spectroscopy acquisition, and MS analysis)
Model organism
Mammalian cell culture (species not specified)
Biosafety
BSL-1

Steps

1
Culture and prepare cells for drug treatment

Establish mammalian cell cultures using standard aseptic techniques and maintain appropriate cell density and viability prior to drug exposure.

▶ 01:06
2
Expose cells to drug compounds

Treat cultured cells with pharmaceutical compounds at defined concentrations and incubation times to allow drug uptake and metabolism.

▶ 01:44
3
Acquire Raman spectral images and process data

Perform confocal Raman spectroscopy on cells to collect spectral signatures of cellular biochemical composition, then apply spectral processing algorithms to extract meaningful metabolic information.

▶ 02:33
4
Sample single cells and perform mass spectrometry

Isolate individual cells using microfluidic or capillary sampling methods and analyze their contents via tandem MS to quantify drug molecules and metabolites.

▶ 03:29
5
Analyze and compare drug-treated versus control cells

Compare Raman and MS data between drug-exposed and untreated cells to identify metabolic changes and drug accumulation patterns at single-cell resolution.

▶ 05:34
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