Effect of laser irradiation on cell function and its implications in Raman spectroscopy

Yuan, X. et al. (2018) Effect of laser irradiation on cell function and its implications in Raman spectroscopy. Applied and Environmental Microbiology, (doi:10.1128/AEM.02508-17) (Early Online Publication)

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Abstract

Lasers are instrumental in advanced bioimaging and Raman spectroscopy. However, they are also well known for their destructive effects on living organisms, leading to concerns about the adverse effects of laser technologies. To implement Raman spectroscopy for cell analysis and manipulation, such as Raman activated cell sorting, it is crucial to identify non-destructive conditions for living cells. Here, we evaluated quantitatively the effect of 532 nm laser irradiation on bacterial cell fate and growth at the single-cell level. Using a purpose-built microfluidic platform, we were able to quantify the growth characteristics i.e. specific growth rate and lag time of individual cells as well as the survival rate of a population in conjunction with Raman spectroscopy. Representative Gram-negative and Gram-positive species show a similar trend in response to laser irradiation dose. Laser irradiation could compromise physiological function of cells and the degree of destruction is both dose and strain dependent, ranging from reduced cell growth to a complete loss of cell metabolic activity and finally to physical disintegration. Gram-positive bacterial cells are more susceptible than Gram-negative bacterial strains to irradiation-induced damage. By directly correlating Raman acquisition with single cell growth characteristics, we provide evidence of non-destructive characteristics of Raman spectroscopy on individual bacterial cells. However, while strong Raman signals can be obtained without causing cell death, the variety of responses from different strains and from individual cells justify careful evaluation of Raman acquisition conditions if cell viability is critical.

Item Type:Articles
Status:Early Online Publication
Refereed:Yes
Glasgow Author(s) Enlighten ID:Yin, Professor Huabing and Cooper, Professor Jonathan and Yuan, Dr Xiaofei and Song, Ms Yanqing and Glidle, Dr Andrew and Ijaz, Dr Umer Zeeshan
Authors: Yuan, X., Song, Y., Song, Y., Xu, J., Wu, Y., Glidle, A., Cusack, M., Ijaz, U. Z., Cooper, J. M., Huang, W. E., and Yin, H.
College/School:College of Science and Engineering > School of Engineering > Biomedical Engineering
College of Science and Engineering > School of Engineering > Infrastructure and Environment
Journal Name:Applied and Environmental Microbiology
Publisher:American Society for Microbiology.
ISSN:0099-2240
ISSN (Online):1098-5336
Published Online:02 February 2018

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Project CodeAward NoProject NamePrincipal InvestigatorFunder's NameFunder RefLead Dept
731921Stable Isotope Probing with Resonance Raman Cell Sorting to profile influence of ocean acidification on microbial carbon fixationHuabing YinNatural Environment Research Council (NERC)NE/P003826/1ENG - BIOMEDICAL ENGINEERING
652771Understanding microbial community through in situ environmental 'omic data synthesisUmer Zeeshan IjazNatural Environment Research Council (NERC)NE/L011956/1ENG - ENGINEERING INFRASTRUCTURE & ENVIR