The sample preparation for cryo-SEM: the real ultrastructure of microbial biofilm or just artifacts?

Result code in IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F68081731%3A_____%2F16%3A00465339" target="_blank" >RIV/68081731:_____/16:00465339 - isvavai.cz</a>

Alternative codes found

RIV/60077344:_____/16:00465339

Result on the web

<a href="http://dx.doi.org/10.1002/9783527808465.EMC2016.6907" target="_blank" >http://dx.doi.org/10.1002/9783527808465.EMC2016.6907</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1002/9783527808465.EMC2016.6907" target="_blank" >10.1002/9783527808465.EMC2016.6907</a>

Result language

angličtina

Original language name

The sample preparation for cryo-SEM: the real ultrastructure of microbial biofilm or just artifacts?

Original language description

The cryo-scanning electron microscopy (cryo-SEM) belongs to reputable techniques in electron microscopy of hydrated samples such as biofilms. The crucial steps of the cryo-preparation techniques are primarily the cryo-fixation and partial sublimation of ice contamination caused during the transfer of the sample to the cryo-high-vacuum preparation chamber where the sublimation process is performed; optionally the freeze-fracturing or coating by metal sputtering or carbon evaporation can be applied. In the case of cryo-fixation, an effort is to keep the frozen biofilm in the form nearby its native state. One of the simplest cryo-fixation techniques is a plunging of the biofilm on a substrate into a liquid cryogen. However, the plunging into a liquid nitrogen or even liquid ethane/propane is sufficient for fixation of very thin layers of biofilm (no more than a few micrometers in thickness) because it is very difficult to achieve enough cooling rates to produce amorphous ice in the sample due to the Leidenfrost effect. Moreover, we show that the cryo-fixation into liquid nitrogen can lead to significant lateral macro-segregation of both bacteria and extracellular polymeric substances (EPS), where plunging into liquid ethane leads to micro-segregation of EPS and macro-segregation of bacteria. Substantially more effective cooling can be achieved by increasing the pressure during exposure to the liquid cryogen. This can be performed for example by the high-pressure freezing (HPF) technique. It was proved that cryo-fixed biofilms by HPF show significantly improved preservation of bacterial ultrastructure and biofilm organization.

Czech name

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Czech description

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Type

D - Article in proceedings

CEP classification

JA - Electronics and optoelectronics

OECD FORD branch

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Project

Result was created during the realization of more than one project. More information in the Projects tab.

Continuities

I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Publication year

2016

Confidentiality

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

Article name in the collection

EMC2016. The 16th European Microscopy Congress. Proceedings

ISBN

9783527808465

ISSN

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e-ISSN

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Number of pages

2

Pages from-to

203-204

Publisher name

Wiley

Place of publication

Oxford

Event location

Lyon

Event date

Aug 28, 2016

Type of event by nationality

WRD - Celosvětová akce

UT code for WoS article

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