Flow Cytometry in Microbiology: The Reason and the Need
The diagnosis of infection is based on methodologies that are, even in the 21st century, based on the study of the ability of microorganisms to grow in the presence of different substrates in the case of identification or in the presence of different anti
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tract The diagnosis of infection is based on methodologies that are, even in the 21st century, based on the study of the ability of microorganisms to grow in the presence of different substrates in the case of identification or in the presence of different antimicrobial drugs in the case of susceptibility evaluation. Despite the use of revolutionary techniques like molecular biology and more recently mass spectrometry, a lot of effort needs to be made to speed the results and to understand what happens to cells as individuals and not just as populations. Flow cytometry is an excellent tool still unexplored in microbiology; several applications are here described in the hope of contributing to the real use of flow cytometry in the clinical lab and to inspire future applications.
Keywords Flow cytometry Identification of microorganisms Susceptibility phenotype determination Mechanisms of resistance Drug monitoring Microbial adhesion Bacteria Fungi Parasites
C. Pina-Vaz (&) S. Costa-de-Oliveira A. Silva-Dias A.P. Silva R. Teixeira-Santos A.G. Rodrigues Department of Microbiology, University of Porto and Faculty of Medicine, Al. Hernâni Monteiro, 4200-319 Porto, Portugal e-mail: [email protected]; [email protected] S. Costa-de-Oliveira e-mail: [email protected] A. Silva-Dias e-mail: [email protected] A.P. Silva e-mail: [email protected] R. Teixeira-Santos e-mail: [email protected] A.G. Rodrigues e-mail: [email protected] © Springer Nature Singapore Pte Ltd. 2017 J.P. Robinson and A. Cossarizza (eds.), Single Cell Analysis, Series in BioEngineering, DOI 10.1007/978-981-10-4499-1_7
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1 Introduction Classic microbiological diagnosis is commonly based upon the study of microbial ability to use different substrates during the growth process for identification and on the measurement of growth inhibition in the presence of different antimicrobials for definition of the antimicrobial susceptibility phenotype. Such cornerstones are invariably time-consuming procedures since the microorganisms need time to replicate. Developments in molecular biology and in immunology, namely monoclonal antibodies and more recently mass spectrometry, have provided new tools to microbiologists, particularly with respect to identification strategies, but not in the area of antimicrobial susceptibility profiles. Flow cytometry (FC) has a strong potential to change the diagnostic paradigm in microbiology. Individual cells can be studied regarding their morphology or structure, or even more interesting, regarding functional aspects. Kinetic studies can be performed, evaluating cell events over time. The potential to sort different microbial cell populations represent a challenge almost completely unexplored. The ancestors of modern flow cytometers were apparatus used during World War II by the US Army in experiments for the detection of bacteria and spores [1]. Many technical improvements were achieved before publication of Flow Cytometry in Microbiology from which most microbiological cyto
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