Monitoring of Multilayered Bacterial Biofilm Morphology by Cryo-SEM for Raman Spectroscopy Measurements

0450101 - ÚPT 2016 RIV US eng J - Journal Article
Hrubanová, Kamila - Bernatová, Silvie - Samek, Ota - Šerý, Mojmír - Zemánek, Pavel - Nebesářová, Jana - Růžička, F. - Krzyžánek, Vladislav
Monitoring of Multilayered Bacterial Biofilm Morphology by Cryo-SEM for Raman Spectroscopy Measurements.
Microscopy and Microanalysis. Roč. 21, S3 (2015), s. 187-188. ISSN 1431-9276. E-ISSN 1435-8115
R&D Projects: GA MŠMT(CZ) LO1212; GA MŠMT ED0017/01/01; GA ČR(CZ) GA14-20012S
Institutional support: RVO:68081731 ; RVO:60077344
Keywords : multilayered bacterial biofilm * morphology by Cryo-SEM * Raman spectroscopy
Subject RIV: JA - Electronics ; Optoelectronics, Electrical Engineering
Impact factor: 1.730, year: 2015

Staphylococcus epidermidis has been recently recognized as an important cause of serious biofilm infections associated with implanted medical devices. In presented work the multi-layered biofilms formed by these microorganisms were observed by scanning electron microscope (SEM), in particular with using freeze-fracturing technique. The freeze-fracture technique consists of physically breaking apart (fracturing) a rapidly frozen biological sample; structural detail exposed by the fracture plane is then visualized by metal deposition. An optional step, involving vacuum sublimation of ice, may be carried out after fracturing. Freeze fracture is unique among electron microscopic techniques in providing planar views of the internal organization of membranes or biofilms. Deep etching of ultra-rapidly frozen samples permits visualization of the surface structure of cells and their components. Our sample was fractured after rapid-freezing into liquid nitrogen, the air-water was removed by short freeze-drying at Alto 2500 chamber (Gatan), sputtered by 5 nm Pt-Pd and finally imaged at low temperature in the field emission SEM JSM 7401F (JEOL).
Permanent Link: http://hdl.handle.net/11104/0251506