Continuous cell sorting in a flow based on single cell resonance Raman spectra

McIlvanna, D., Huang, W. E., Davison, P., Glidle, A., Cooper, J. and Yin, H. (2016) Continuous cell sorting in a flow based on single cell resonance Raman spectra. Lab on a Chip, 16(8), pp. 1420-1429. (doi:10.1039/C6LC00251J) (PMID:26974400)

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Abstract

Single cell Raman spectroscopy measures a spectral fingerprint of the biochemistry of cells, and provides a powerful method for label-free detection of living cells without the involvement of a chemical labelling strategy. However, as the intrinsic Raman signals of cells are inherently weak, there is a significant challenge in discriminating and isolating cells in a flowing stream. Here we report an integrated Raman-microfluidic system for continuous sorting of a stream of cyanobacteria, Synechocystis sp. PCC6803. These carotenoidcontaining microorganisms provide an elegant model system enabling us to determine the sorting accuracy using the subtly different resonance Raman spectra of microorganism cultured in a 12C or 13C carbon source. Central to the implementation of continuous flow sorting is the use of “pressure dividers” that eliminate fluctuations in flow in the detection region. This has enabled us to stabilise the flow profile sufficiently to allow automated operation with synchronisation of Raman acquisition, real-time classification and sorting at flow rates of ca. <100 μm/s, without the need to “trap” the cells. We demonstrate the flexibility of this approach in sorting mixed cell populations with the ability to achieve 96.3% purity of the selected cells at a speed of 0.5 Hz.

Item Type:Articles
Additional Information:The authors thank partial support from EPSRC (EP/H04986X/1 and EP/J009121/1). We gratefully acknowledge the technical team of the James Watt Nanofabrication Centre (JWNC) at University of Glasgow for the support in fabricating the devices.
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Yin, Professor Huabing and Cooper, Professor Jonathan and Glidle, Dr Andrew
Authors: McIlvanna, D., Huang, W. E., Davison, P., Glidle, A., Cooper, J., and Yin, H.
College/School:College of Science and Engineering > School of Engineering > Biomedical Engineering
Journal Name:Lab on a Chip
Publisher:Royal Society of Chemistry
ISSN:1473-0197
ISSN (Online):1473-0189
Published Online:02 March 2016
Copyright Holders:Copyright © 2016 Royal Society of Chemistry
First Published:First published in Lab on a Chip 16(8):1420-1429
Publisher Policy:Reproduced under a Creative Commons License
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Project CodeAward NoProject NamePrincipal InvestigatorFunder's NameFunder RefLead Dept
532401Engineering a semi-biotic systemHuabing YinEngineering & Physical Sciences Research Council (EPSRC)EP/H04986X/1ENG - BIOMEDICAL ENGINEERING
574961Shell inspiration: turning nature's secrets into engineering solutions.Huabing YinEngineering & Physical Sciences Research Council (EPSRC)EP/J009121/1ENG - BIOMEDICAL ENGINEERING