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Non-contact, Label-free Monitoring of Cells and Extracellular Matrix using Raman Spectroscopy
Published on: May 29, 2012
Monitoring cellular behaviour using Raman spectroscopy for tissue engineering and regenerative medicine applications
A R Boyd1, G A Burke, B J Meenan
1School of Engineering, Nanotechnology, University of Ulster at Jordanstown, Newtownabbey, Co. Antrim, Northern Ireland, UK. ar.boyd@ulster.ac.uk
Journal of Materials Science. Materials in Medicine
|December 19, 2009
Summary
Raman spectroscopy can rapidly assess live cells non-invasively for tissue engineering and regenerative medicine (TERM). While unable to distinguish cell lines, it effectively differentiates viable from non-viable cells by detecting biochemical changes post-death.
Area of Science:
- Biophotonics
- Spectroscopy
- Biomaterials Science
Background:
- Raman spectroscopy offers chemical composition analysis, experiencing a resurgence due to technological advancements.
- Traditional in vitro cell analysis methods are time-consuming and can introduce artifacts.
- Tissue Engineering and Regenerative Medicine (TERM) requires efficient, non-invasive cell monitoring.
Purpose of the Study:
- To evaluate a macro-sampling Raman spectrometer for rapid, non-invasive analysis of TERM-relevant cells.
- To determine if Raman spectroscopy can discriminate between different cell types and assess cell viability.
- To explore the potential of Raman spectroscopy in quality assurance frameworks for TERM.
Main Methods:
- Utilized a versatile Raman spectrometer with a macro-sampling optical platform (100 µm laser spot size).
- Acquired Raman spectra from live TERM-relevant cells in a non-destructive manner.
- Analyzed spectral differences between viable and non-viable cells, focusing on protein, DNA/RNA, and lipid constituents.
Main Results:
- Demonstrated the capability of obtaining rapid, non-invasive Raman spectra from live cells.
- Observed significant spectral differences between viable and non-viable cells, indicating changes in biochemical composition upon cell death.
- Was not able to discriminate between different cell lines in this study.
Conclusions:
- Macro-sampling Raman spectroscopy shows significant potential for non-destructive, real-time analysis in TERM.
- The technique can identify cell death by detecting biochemical alterations.
- This method offers a pathway for quality assurance in TERM by providing direct data from live cells.
Related Concept Videos
01:20Raman 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...
However, a small fraction of the scattered light exhibits a frequency shift due to the exchange of energy between the incident photons and the...
01:26Raman 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...
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...