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Non-invasive analysis of stored red blood cells using diffuse resonance Raman spectroscopy
Rekha Gautam1, Joo-Yeun Oh, Rakesh P Patel
1Department of Chemistry, University of Alabama at Birmingham, Birmingham, AL 35294, USA. rdluhy@uab.edu.
The Analyst
|July 24, 2018
Summary
A new diffuse resonance Raman spectroscopy (DRRS) method non-invasively analyzes sub-surface components like red blood cells (RBCs). This technique successfully differentiated young from old RBCs, showing promise for assessing biochemical changes during storage.
Area of Science:
- Spectroscopy
- Biomedical Optics
- Biochemistry
Background:
- Biochemical changes occur in stored red blood cells (RBCs), known as the "storage lesion."
- Current non-invasive methods to assess RBC storage lesion are lacking.
- Sub-surface component analysis is challenging in biological samples.
Purpose of the Study:
- To develop and validate a novel non-invasive method for acquiring Raman spectra of sub-surface components.
- To apply this method for analyzing biochemical changes in stored red blood cells (RBCs).
- To assess the potential of the technique for differentiating RBCs based on storage age.
Main Methods:
- Utilized diffusely focused radiation in a microscope sampling configuration to generate Raman scattering at various depths.
- Employed defocused laser illumination and increased CCD pixel binning for diffuse sampling.
- Applied diffuse resonance Raman spectroscopy (DRRS) to analyze RBCs through a 1 mm blood bag wall and compared it with stand-off Raman spectroscopy (SORS).
Main Results:
- Optimized instrumental parameters (pixel binning, slit width, bandwidth) for maximizing diffusely scattered signals.
- Successfully detected RBCs through a blood bag segment using DRRS.
- Differentiated young (6-8 day) from old (35-40 day) RBCs with 100% sensitivity and 98.5% selectivity using time-dependent DRRS spectra and a multivariate model.
Conclusions:
- DRRS is a promising non-invasive technique for sub-surface component spectral acquisition.
- The method is particularly effective for samples exhibiting resonant enhancement.
- DRRS shows significant potential for monitoring biochemical changes in stored RBCs and other biological samples.
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