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Related Concept Videos

Raman Spectroscopy Instrumentation: Overview01:26

Raman Spectroscopy Instrumentation: Overview

A conventional Raman spectrophotometer includes a laser source, a sample holding system, a wavelength selector, and a detector.
The monochromatic laser source, typically using visible or near-infrared radiation, generates a highly focused beam of light. This light interacts with the molecules of the sample, scattering some of the light. Liquid and gaseous samples are usually tested in ordinary glass capillaries, while solids can be analyzed as powders packed in capillaries or as potassium...
Raman Spectroscopy: Overview01:20

Raman Spectroscopy: Overview

The underlying principle of Raman spectroscopy is based on the interaction between light and matter, specifically molecules' inelastic scattering of photons. When a monochromatic beam of light, typically from a laser source, interacts with a sample, most scattered light has the same frequency as the incident light. This is known as Rayleigh scattering.
However, a small fraction of the scattered light exhibits a frequency shift due to the exchange of energy between the incident photons and the...

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Surface-enhanced Resonance Raman Scattering Nanoprobe Ratiometry for Detecting Microscopic Ovarian Cancer via Folate Receptor Targeting
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Identifying and localizing intracellular nanoparticles using Raman spectroscopy.

Jennifer Dorney1, Franck Bonnier, Amaya Garcia

  • 1Nanolab Research Centre, Focas Research Institute, Dublin Institute of Technology (DIT), Camden Row, Dublin, 8, Ireland.

The Analyst
|January 26, 2012
PubMed
Summary

Raman microscopy successfully located polystyrene nanoparticles within cells, revealing their endoplasmic reticulum localization and lipid-rich environment. This technique offers a non-toxic method for tracking nanoparticles and understanding cellular responses.

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Area of Science:

  • Spectroscopy and microscopy
  • Nanotechnology and cell biology
  • Biophysics

Background:

  • Fluorescently labelled nanoparticles are difficult to track within cells due to diffuse fluorescence.
  • Conventional microscopy struggles to unambiguously identify nanoparticle distribution and subcellular localization.
  • Understanding nanoparticle-cell interactions requires precise localization and environmental analysis.

Purpose of the Study:

  • To develop and validate Raman microscopy combined with K-means clustering and PCA for precise nanoparticle localization within biological cells.
  • To assess the non-toxicity of polystyrene nanoparticles using standard cytotoxicity assays.
  • To identify the subcellular environment and metabolic changes associated with nanoparticle exposure.

Main Methods:

  • Raman spectroscopic mapping of A549 cells exposed to polystyrene nanoparticles.
  • K-means clustering and Principal Component Analysis (PCA) for spectral data analysis.
  • Cytotoxicity assays (Neutral Red, Alamar Blue, Coomassie Blue, MTT) to evaluate nanoparticle toxicity.

Main Results:

  • Raman microscopy with K-means clustering unambiguously localized 50 nm and 100 nm polystyrene nanoparticles within cells.
  • PCA identified nanoparticle localization in lipid-rich regions, suggesting association with the endoplasmic reticulum.
  • Cytotoxicity assays confirmed the non-toxic nature of the nanoparticles to A549 cells.
  • Confocal fluorescence microscopy showed diffuse fluorescence, failing to provide clear localization.

Conclusions:

  • Raman microscopy is a powerful tool for unambiguous identification and localization of nonfluorescent nanoparticles in cells.
  • The technique can probe the cellular environment and metabolic changes related to nanoparticle interactions.
  • Optimized cell preparation is crucial for successful Raman spectroscopic analysis of nanoparticle-cell interactions.