Assembly for Cryo-SEM/Raman Microspectroscopy sample analysis

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F68081731%3A_____%2F22%3A00568569" target="_blank" >RIV/68081731:_____/22:00568569 - isvavai.cz</a>

Výsledek na webu

<a href="https://www.16mcm.cz/wp-content/uploads/2022/09/16MCM-abstract-book.pdf" target="_blank" >https://www.16mcm.cz/wp-content/uploads/2022/09/16MCM-abstract-book.pdf</a>

DOI - Digital Object Identifier

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Jazyk výsledku

angličtina

Název v původním jazyce

Assembly for Cryo-SEM/Raman Microspectroscopy sample analysis

Popis výsledku v původním jazyce

Correlative imaging using different methods of observation is now becoming more and more popular. It allows the interconnection of various information of the sample accessible only bynindividual techniques. It is crucial to find and observe the same place on the sample. The same objects and their locations (cell shape and arrangement) can be correlated with each other,nespecially with software when 2 different imaging devices were used. Combination of the Cryo-Scanning Electron Microscopy (Cryo-SEM) and Cryo-Raman Micro-spectroscopy (Cryo-Raman) gives many benefits. Surface, structure or morphology of the sample can by studied using Cryo-SEM, and detection and identification of elements or compounds in the examined sample using Cryo-Raman. For observation of various, especially biological samples, their freezing to the temperature of liquid nitrogen is used. The motivation was to design an assembly for motion and observation using Cryo-Raman and its connection with commercial holders used for the observation of vitrified samples by Cryo-SEM. Assembly allows movement of the sample to a known position and measuring the cryogen level using LabVIEW software. Our assembly consists of a Dewar vessel, which contains liquid nitrogen. The stage is inside the Dewar vessel below the liquid level and is made by 3D printing and enables 3 positions for individual types of sample holders (Leica microsystems, Gatan and Quorum technologies). The Dewar vessel lies on piezoelectric stage allowing movement in both X and Y direction. The stage is connected to the Raman spectrometer table providing displacement in the Z direction. The Dewar vessel is covered with Plexiglas to minimize contamination. Assembly was tested on dried samples and polystyrene in cryo temperatures with known spectrum to verify functionality. Then biological samples including C. necator H16 were tested. System allows more efficient correlation between individual analyses of microorganisms containing biopolymer particles. For example, several representatives of soil microorganisms, such as bacteria C. necator H16 which are able to produce polyhydroxyalkanoates (PHA). PHAs are polyesters of hydroxyalkanoic acids, which serve as energy storage. PHAs attract attention as an alternative to petrochemical plastics, which can be biotechnologically produced from waste streams of some industrial processes. It was proved that the combination of Raman spectroscopy with the cryo-SEM technique can provide a deeper insight into the chemical and mechanical properties of polymeric granules inside the bacterial cells and it will serve for future research.

Název v anglickém jazyce

Assembly for Cryo-SEM/Raman Microspectroscopy sample analysis

Popis výsledku anglicky

Correlative imaging using different methods of observation is now becoming more and more popular. It allows the interconnection of various information of the sample accessible only bynindividual techniques. It is crucial to find and observe the same place on the sample. The same objects and their locations (cell shape and arrangement) can be correlated with each other,nespecially with software when 2 different imaging devices were used. Combination of the Cryo-Scanning Electron Microscopy (Cryo-SEM) and Cryo-Raman Micro-spectroscopy (Cryo-Raman) gives many benefits. Surface, structure or morphology of the sample can by studied using Cryo-SEM, and detection and identification of elements or compounds in the examined sample using Cryo-Raman. For observation of various, especially biological samples, their freezing to the temperature of liquid nitrogen is used. The motivation was to design an assembly for motion and observation using Cryo-Raman and its connection with commercial holders used for the observation of vitrified samples by Cryo-SEM. Assembly allows movement of the sample to a known position and measuring the cryogen level using LabVIEW software. Our assembly consists of a Dewar vessel, which contains liquid nitrogen. The stage is inside the Dewar vessel below the liquid level and is made by 3D printing and enables 3 positions for individual types of sample holders (Leica microsystems, Gatan and Quorum technologies). The Dewar vessel lies on piezoelectric stage allowing movement in both X and Y direction. The stage is connected to the Raman spectrometer table providing displacement in the Z direction. The Dewar vessel is covered with Plexiglas to minimize contamination. Assembly was tested on dried samples and polystyrene in cryo temperatures with known spectrum to verify functionality. Then biological samples including C. necator H16 were tested. System allows more efficient correlation between individual analyses of microorganisms containing biopolymer particles. For example, several representatives of soil microorganisms, such as bacteria C. necator H16 which are able to produce polyhydroxyalkanoates (PHA). PHAs are polyesters of hydroxyalkanoic acids, which serve as energy storage. PHAs attract attention as an alternative to petrochemical plastics, which can be biotechnologically produced from waste streams of some industrial processes. It was proved that the combination of Raman spectroscopy with the cryo-SEM technique can provide a deeper insight into the chemical and mechanical properties of polymeric granules inside the bacterial cells and it will serve for future research.

Druh

O - Ostatní výsledky

CEP obor

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OECD FORD obor

20201 - Electrical and electronic engineering

Projekt

<a href="/cs/project/TN01000008" target="_blank" >TN01000008: Centrum elektronové a fotonové optiky</a><br>

Návaznosti

I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Rok uplatnění

2022

Kód důvěrnosti údajů

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