The innovation of cryo-SEM freeze-fracturing methodology
demonstrated on high pressure frozen biofilm

J 2018

The innovation of cryo-SEM freeze-fracturing methodology demonstrated on high pressure frozen biofilm

HRUBANOVA, Kamila, Jana NEBESAROVA, Filip RŮŽIČKA a Vladislav KRZYZANEK

Základní údaje

Originální název

The innovation of cryo-SEM freeze-fracturing methodology demonstrated on high pressure frozen biofilm

Autoři

HRUBANOVA, Kamila (203 Česká republika), Jana NEBESAROVA (203 Česká republika), Filip RŮŽIČKA (203 Česká republika, domácí) a Vladislav KRZYZANEK (203 Česká republika, garant)

Vydání

Micron, OXFORD, PERGAMON-ELSEVIER SCIENCE LTD, 2018, 0968-4328

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Obor

10606 Microbiology

Stát vydavatele

Velká Británie a Severní Irsko

Utajení

není předmětem státního či obchodního tajemství

Impakt faktor

Impact factor: 1.530

Kód RIV

RIV/00216224:14110/18:00104166

Organizační jednotka

Lékařská fakulta

DOI

http://dx.doi.org/10.1016/j.micron.2018.04.006

UT WoS

000435048000004

Klíčová slova anglicky

cryo-SEM; High-pressure freezing (HPF); Freeze-fracturing; Biofilm; Candida; Staphylococcus

Štítky

14110113, rivok

Příznaky

Mezinárodní význam, Recenzováno

Změněno: 11. 2. 2019 14:10, Soňa Böhmová

Anotace

V originále

In this study we present an innovative method for the preparation of fully hydrated samples of microbial biofilms of cultures Staphylococcus epidermidis, Candida parapsilosis and Candida albicans. Cryo-scanning electron micro-scopy (cryo-SEM) and high-pressure freezing (HPF) rank among cutting edge techniques in the electron microscopy of hydrated samples such as biofilms. However, the combination of these techniques is not always easily applicable. Therefore, we present a method of combining high-pressure freezing using EM PACT2 (Leica Microsystems), which fixes hydrated samples on small sapphire discs, with a high resolution SEM equipped with the widely used cryo-preparation system ALTO 2500 (Gatan). Using a holder developed in house, a freeze fracturing technique was applied to image and investigate microbial cultures cultivated on the sapphire discs. In our experiments, we focused on the ultrastructure of the extracellular matrix produced during cultivation and the relationships among microbial cells in the biofilm. The main goal of our investigations was the detailed visualization of areas of the biofilm where the microbial cells adhere to the substrate/surface. We show the feasibility of this technique, which is clearly demonstrated in experiments with various freeze-etching times.

Návaznosti

NV16-29916A, projekt VaV

Název: Využití bakteriofágů v léčbě nozokomiálních infekcí spojených s multirezistencí či tvorbou biofilmu

Citovat

HRUBANOVA, Kamila, Jana NEBESAROVA, Filip RŮŽIČKA a Vladislav KRZYZANEK. The innovation of cryo-SEM freeze-fracturing methodology demonstrated on high pressure frozen biofilm. Micron. OXFORD: PERGAMON-ELSEVIER SCIENCE LTD, 2018, roč. 110, JUL 2018, s. 28-35. ISSN 0968-4328. Dostupné z: https://dx.doi.org/10.1016/j.micron.2018.04.006.

@article{1454496,
   author = {Hrubanova, Kamila and Nebesarova, Jana and Růžička, Filip and Krzyzanek, Vladislav},
   article_location = {OXFORD},
   article_number = {JUL 2018},
   doi = {http://dx.doi.org/10.1016/j.micron.2018.04.006},
   keywords = {cryo-SEM; High-pressure freezing (HPF); Freeze-fracturing; Biofilm; Candida; Staphylococcus},
   language = {eng},
   issn = {0968-4328},
   journal = {Micron},
   title = {The innovation of cryo-SEM freeze-fracturing methodology demonstrated on high pressure frozen biofilm},
   volume = {110},
   year = {2018}
}

TY  - JOUR
ID  - 1454496
AU  - Hrubanova, Kamila - Nebesarova, Jana - Růžička, Filip - Krzyzanek, Vladislav
PY  - 2018
TI  - The innovation of cryo-SEM freeze-fracturing methodology demonstrated on high pressure frozen biofilm
JF  - Micron
VL  - 110
IS  - JUL 2018
SP  - 28-35
EP  - 28-35
PB  - PERGAMON-ELSEVIER SCIENCE LTD
SN  - 09684328
KW  - cryo-SEM
KW  - High-pressure freezing (HPF)
KW  - Freeze-fracturing
KW  - Biofilm
KW  - Candida
KW  - Staphylococcus
N2  - In this study we present an innovative method for the preparation of fully hydrated samples of microbial biofilms of cultures Staphylococcus epidermidis, Candida parapsilosis and Candida albicans. Cryo-scanning electron micro-scopy (cryo-SEM) and high-pressure freezing (HPF) rank among cutting edge techniques in the electron microscopy of hydrated samples such as biofilms. However, the combination of these techniques is not always easily applicable. Therefore, we present a method of combining high-pressure freezing using EM PACT2 (Leica Microsystems), which fixes hydrated samples on small sapphire discs, with a high resolution SEM equipped with the widely used cryo-preparation system ALTO 2500 (Gatan). Using a holder developed in house, a freeze fracturing technique was applied to image and investigate microbial cultures cultivated on the sapphire discs. In our experiments, we focused on the ultrastructure of the extracellular matrix produced during cultivation and the relationships among microbial cells in the biofilm. The main goal of our investigations was the detailed visualization of areas of the biofilm where the microbial cells adhere to the substrate/surface. We show the feasibility of this technique, which is clearly demonstrated in experiments with various freeze-etching times.
ER  -

HRUBANOVA, Kamila, Jana NEBESAROVA, Filip RŮŽIČKA a Vladislav KRZYZANEK. The innovation of cryo-SEM freeze-fracturing methodology demonstrated on high pressure frozen biofilm. \textit{Micron}. OXFORD: PERGAMON-ELSEVIER SCIENCE LTD, 2018, roč.~110, JUL 2018, s.~28-35. ISSN~0968-4328. Dostupné z: https://dx.doi.org/10.1016/j.micron.2018.04.006.

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