Optimization of confocal microscopy for visualization and quantification of yeast biofilm

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F26722861%3A_____%2F18%3AN0000079" target="_blank" >RIV/26722861:_____/18:N0000079 - isvavai.cz</a>

Výsledek na webu

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DOI - Digital Object Identifier

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Jazyk výsledku

angličtina

Název v původním jazyce

Optimization of confocal microscopy for visualization and quantification of yeast biofilm

Popis výsledku v původním jazyce

Poster a sborník abstrakt: Smolová, J., Němečková, I., Turoňová, H.: Optimization of confocal microscopy for visualization and quantification of yeast biofilm. Poster. 5th World Congress on Microbial Biotechnology, 17. -18. 9. 2018, Lisbon, Portugalsko. Sborník abstrakt str. 36. Due to the resistance of biofilm to conventional cleaning processes, an industrial biofilm, especially in dairy industry, could cause extensive problems in final products in case of insufficient sanitation process. Our work focuses on visualization and quantification of yeasts biofilm using confocal microscopy. Commercial dying kits are optimized for pathogenic bacteria, therefore, the dying protocol for yeasts must have been created. The yeasts were isolated as industrial contaminants from cheese and saline solution and were identified as Yarrowia lipolytica and Debaryomyces hansenii using MALDI-TOF. Material and methods: µCLEAR® Chimney well plate was used for yeasts cultivation and the cultivation time, washing, dying and visualization of yeasts biofilm was optimized. LIVE/DEAD® BacLightTM Bacterial Viability and Counting Kit was used for dying and the biofilm visualization was performed by a confocal laser scanning microscopy with a rotating disc, a ten-fold magnification and a 1 μm z-step. For the living, dead and damaged cells, biomass volume and total biofilm structure were determined using Imaris software. Results: The cultivation time was set to 24 hours with fresh broth well washing after 2 hours which better corresponds with real conditions. While living cells predominated in D. hansenii biofilm, the damaged cells prevailed in the Y. lipolytica biofilm, which may have been related to the different aging rates of the biofilm of both strains. The dying time was shortened to 60 minutes to reduce the toxic effect of propidium iodide. Conclusion: New approaches for visualization and quantification of biofilm are needed for better understanding of the biofilm forming and persistence. We have shown that visualization of yeast biofilm is one of the ways to study these biofilms. These findings can lead to monitoring the effectiveness of sanitation solutions and to designing better cleaning processes.

Název v anglickém jazyce

Optimization of confocal microscopy for visualization and quantification of yeast biofilm

Popis výsledku anglicky

Poster a sborník abstrakt: Smolová, J., Němečková, I., Turoňová, H.: Optimization of confocal microscopy for visualization and quantification of yeast biofilm. Poster. 5th World Congress on Microbial Biotechnology, 17. -18. 9. 2018, Lisbon, Portugalsko. Sborník abstrakt str. 36. Due to the resistance of biofilm to conventional cleaning processes, an industrial biofilm, especially in dairy industry, could cause extensive problems in final products in case of insufficient sanitation process. Our work focuses on visualization and quantification of yeasts biofilm using confocal microscopy. Commercial dying kits are optimized for pathogenic bacteria, therefore, the dying protocol for yeasts must have been created. The yeasts were isolated as industrial contaminants from cheese and saline solution and were identified as Yarrowia lipolytica and Debaryomyces hansenii using MALDI-TOF. Material and methods: µCLEAR® Chimney well plate was used for yeasts cultivation and the cultivation time, washing, dying and visualization of yeasts biofilm was optimized. LIVE/DEAD® BacLightTM Bacterial Viability and Counting Kit was used for dying and the biofilm visualization was performed by a confocal laser scanning microscopy with a rotating disc, a ten-fold magnification and a 1 μm z-step. For the living, dead and damaged cells, biomass volume and total biofilm structure were determined using Imaris software. Results: The cultivation time was set to 24 hours with fresh broth well washing after 2 hours which better corresponds with real conditions. While living cells predominated in D. hansenii biofilm, the damaged cells prevailed in the Y. lipolytica biofilm, which may have been related to the different aging rates of the biofilm of both strains. The dying time was shortened to 60 minutes to reduce the toxic effect of propidium iodide. Conclusion: New approaches for visualization and quantification of biofilm are needed for better understanding of the biofilm forming and persistence. We have shown that visualization of yeast biofilm is one of the ways to study these biofilms. These findings can lead to monitoring the effectiveness of sanitation solutions and to designing better cleaning processes.

Druh

O - Ostatní výsledky

CEP obor

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OECD FORD obor

21101 - Food and beverages

Projekt

<a href="/cs/project/QK1710156" target="_blank" >QK1710156: Nové přístupy a metody analýzy pro zajištění kvality, bezpečnosti a zdravotní nezávadnosti sýrů, optimalizace jejich výroby a zefektivnění procesů hygieny a sanitace při současném snížení zátěže životního prostředí odpadními vodami</a><br>

Návaznosti

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)<br>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