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Monitoring Candida parapsilosis and Staphylococcus epidermidis Biofilms by a Combination of Scanning Electron Microscopy and Raman Spectroscopy

1
Institute of Scientific Instruments of the Czech Academy of Sciences, CZ-61264 Brno, Czech Republic
2
Biology Centre of the Czech Academy of Sciences, CZ-37005 Ceske Budejovice, Czech Republic
3
Department of Microbiology, Faculty of Medicine, Masaryk University and St. Anne’s Faculty Hospital, CZ-65691 Brno, Czech Republic
*
Author to whom correspondence should be addressed.
Sensors 2018, 18(12), 4089; https://doi.org/10.3390/s18124089
Received: 22 September 2018 / Revised: 16 November 2018 / Accepted: 20 November 2018 / Published: 22 November 2018
(This article belongs to the Special Issue Applications of Raman Spectroscopy in Sensors)
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Abstract

The biofilm-forming microbial species Candida parapsilosis and Staphylococcus epidermidis have been recently linked to serious infections associated with implanted medical devices. We studied microbial biofilms by high resolution scanning electron microscopy (SEM), which allowed us to visualize the biofilm structure, including the distribution of cells inside the extracellular matrix and the areas of surface adhesion. We compared classical SEM (chemically fixed samples) with cryogenic SEM, which employs physical sample preparation based on plunging the sample into various liquid cryogens, as well as high-pressure freezing (HPF). For imaging the biofilm interior, we applied the freeze-fracture technique. In this study, we show that the different means of sample preparation have a fundamental influence on the observed biofilm structure. We complemented the SEM observations with Raman spectroscopic analysis, which allowed us to assess the time-dependent chemical composition changes of the biofilm in vivo. We identified the individual spectral peaks of the biomolecules present in the biofilm and we employed principal component analysis (PCA) to follow the temporal development of the chemical composition. View Full-Text
Keywords: Raman spectroscopy; biofilm; sample preparation; scanning electron microscopy; cryo-SEM Raman spectroscopy; biofilm; sample preparation; scanning electron microscopy; cryo-SEM
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Hrubanova, K.; Krzyzanek, V.; Nebesarova, J.; Ruzicka, F.; Pilat, Z.; Samek, O. Monitoring Candida parapsilosis and Staphylococcus epidermidis Biofilms by a Combination of Scanning Electron Microscopy and Raman Spectroscopy. Sensors 2018, 18, 4089.

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