Optimization of confocal microscopy for visualization and quantification of yeast biofilm

Result code in 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>

Result on the web

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DOI - Digital Object Identifier

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Result language

angličtina

Original language name

Optimization of confocal microscopy for visualization and quantification of yeast biofilm

Original language description

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.

Czech name

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Czech description

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Type

O - Miscellaneous

CEP classification

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OECD FORD branch

21101 - Food and beverages

Project

<a href="/en/project/QK1710156" target="_blank" >QK1710156: New approaches and methods of analysis to ensure the quality, safety and health wholesomeness of cheeses, the optimization of their production and the streamline of hygiene and sanitation together with the lowering of environmental load by waste water.</a><br>

Continuities

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)<br>I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Publication year

2018

Confidentiality

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