Label-free haemogram using wavelength modulated Raman spectroscopy for identifying immune-cell subset
MetadataShow full item record
Leucocytes in the blood of mammals form a powerful protective system against a wide range of dangerous pathogens. There are several types of immune cells that has specific role in the whole immune system. The number and type of immune cells alter in the disease state and identifying the type of immune cell provides information about a person's state of health. There are several immune cell subsets that are essentially morphologically identical and require external labeling to enable discrimination. Here we demonstrate the feasibility of using Wavelength Modulated Raman Spectroscopy (WMRS) with suitable machine learning algorithms as a label-free method to distinguish between different closely lying immune cell subset. Principal Component Analysis (PCA) was performed on WMRS data from single cells, obtained using confocal Raman microscopy for feature reduction, followed by Support Vector Machine (SVM) for binary discrimination of various cell subset, which yielded an accuracy >85%. The method was successful in discriminating between untouched and unfixed purified populations of CD4+CD3+ and CD8+CD3+ T lymphocyte subsets, and CD56+CD3- natural killer cells with a high degree of specificity. It was also proved sensitive enough to identify unique Raman signatures that allow clear discrimination between dendritic cell subsets, comprising CD303+CD45+ plasmacytoid and CD1c+CD141+ myeloid dendritic cells. The results of this study clearly show that WMRS is highly sensitive and can distinguish between cell types that are morphologically identical.
Ashok , P C , Praveen , B B , Campbell , E C , Dholakia , K & Powis , S J 2014 , Label-free haemogram using wavelength modulated Raman spectroscopy for identifying immune-cell subset . in A Mahadevan-Jansen & W Petrich (eds) , Biomedical Vibrational Spectroscopy VI : Advances in Research and Industry . vol. 8939 , Proceedings of SPIE , vol. 8839 , SPIE . DOI: 10.1117/12.2039836
Biomedical Vibrational Spectroscopy VI
Copyright 2014 Society of Photo-Optical Instrumentation Engineers. One print or electronic copy may be made for personal use only. Systematic reproduction and distribution, duplication of any material in this paper for a fee or for commercial purposes, or modification of the content of the paper are prohibited.
DescriptionThe authors thank the UK EPSRC the CR-UK/EPSRC/MRC/DoH (England) imaging Program and the European Union project FAMOS (FP7 ICT, contract no. 317744) for funding, K.D. is a Royal Society-Wolfson Merit Award Holder.
Items in the St Andrews Research Repository are protected by copyright, with all rights reserved, unless otherwise indicated.