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

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F60076658%3A12310%2F18%3A43897418" target="_blank" >RIV/60076658:12310/18:43897418 - isvavai.cz</a>

Nalezeny alternativní kódy

RIV/60077344:_____/18:00491213 RIV/68081731:_____/18:00491213 RIV/00216224:14110/18:00104166 RIV/00159816:_____/18:00068743

Výsledek na webu

<a href="https://reader.elsevier.com/reader/sd/pii/S0968432818300209?token=A5A4808408926CF767C4AB7CA900E48333FEC796F0EB9F7A014D3109DFF923AC5C5A4625C149BB780C41AA53096E99EA" target="_blank" >https://reader.elsevier.com/reader/sd/pii/S0968432818300209?token=A5A4808408926CF767C4AB7CA900E48333FEC796F0EB9F7A014D3109DFF923AC5C5A4625C149BB780C41AA53096E99EA</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1016/j.micron.2018.04.006" target="_blank" >10.1016/j.micron.2018.04.006</a>

Jazyk výsledku

angličtina

Název v původním jazyce

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

Popis výsledku v původním jazyce

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.

Název v anglickém jazyce

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

Popis výsledku anglicky

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.

Druh

J_imp - Článek v periodiku v databázi Web of Science

CEP obor

—

OECD FORD obor

20201 - Electrical and electronic engineering

Projekt

Výsledek vznikl pri realizaci vícero projektů. Více informací v záložce Projekty.

Návaznosti

I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Rok uplatnění

2018

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název periodika

Micron

ISSN

0968-4328

e-ISSN

—

Svazek periodika

110

Číslo periodika v rámci svazku

JUL 2018

Stát vydavatele periodika

GB - Spojené království Velké Británie a Severního Irska

Počet stran výsledku

8

Strana od-do

28-35

Kód UT WoS článku

000435048000004

EID výsledku v databázi Scopus

2-s2.0-85046133186