A 3D printed Microfluidic Flow-Cell for Microscopy Analysis of In Situ-Grown Biofilms

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A 3D printed Microfluidic Flow-Cell for Microscopy Analysis of In Situ-Grown Biofilms. / Kristensen, Mathilde Frost; Leonhardt, Dirk; Neland, Merethe Louise Bønneland et al.

In: Caries Research, 07.2020.

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@article{e147adc2ac464549aacd669b35150319,
title = "A 3D printed Microfluidic Flow-Cell for Microscopy Analysis of In Situ-Grown Biofilms",
abstract = "Metabolic processes inside dental biofilms are likely to be influencedby the velocity of the thin saliva film in contact with the biofilm.In addition, in situ-grown biofilms are likely to behavedifferently than in vitro biofilms. However, there is a lack of commerciallyavailable flow-cells able to incorporate in situ samplesand adjust the saliva film to a match the oral cavity. Therefore, theaim of this study was to design and 3D-print such a microfluidicflow-cell. As a proof of concept, we studied pH developmentsinside three 96 h biofilms under a stimulated saliva flow. Themicroscope compatible flow-cell consists of an incoming port, outcoming port and a central bottomless impression for the in situsample. Sealing the flow-cell to a coverslip forms a bottom to theflow-cell and creates a defined flow-space between the in situ sampleand the coverslip. The participant grew the in situ biofilms oncustom-made glass slabs fitted into a removable splint. The splintwas worn the entire time except during meals, and was immersedin 10 % sucrose three times a day. Unsterile stimulated saliva and4 % sucrose were used as flow medium, and the pH was analyzedusing the ratiometric dye C-SNARF-4. The results of this pilotstudy showed that after 30 min of static conditions, pH ranged from5.7 to 6.9, with differences both between fields of view (FOVs) andbetween biofilms, confirming the presence of microenvironments.After the onset of flow (5 mm/min), pH rose to neutral orslightly alkaline values in all three biofilms. However, differencesbetween FOVs were still present. In conclusion, incorporatingflow is important when studying pH developments inside biofilms.",
author = "Kristensen, {Mathilde Frost} and Dirk Leonhardt and Neland, {Merethe Louise B{\o}nneland} and Sebastian Schlafer",
year = "2020",
month = jul,
language = "English",
journal = "Caries Research",
issn = "0008-6568",
publisher = "S. Karger AG",
note = "null ; Conference date: 09-07-2020 Through 11-07-2020",
url = "https://www.orca-caries-research.org/events/past-events",

}

RIS

TY - ABST

T1 - A 3D printed Microfluidic Flow-Cell for Microscopy Analysis of In Situ-Grown Biofilms

AU - Kristensen, Mathilde Frost

AU - Leonhardt, Dirk

AU - Neland, Merethe Louise Bønneland

AU - Schlafer, Sebastian

PY - 2020/7

Y1 - 2020/7

N2 - Metabolic processes inside dental biofilms are likely to be influencedby the velocity of the thin saliva film in contact with the biofilm.In addition, in situ-grown biofilms are likely to behavedifferently than in vitro biofilms. However, there is a lack of commerciallyavailable flow-cells able to incorporate in situ samplesand adjust the saliva film to a match the oral cavity. Therefore, theaim of this study was to design and 3D-print such a microfluidicflow-cell. As a proof of concept, we studied pH developmentsinside three 96 h biofilms under a stimulated saliva flow. Themicroscope compatible flow-cell consists of an incoming port, outcoming port and a central bottomless impression for the in situsample. Sealing the flow-cell to a coverslip forms a bottom to theflow-cell and creates a defined flow-space between the in situ sampleand the coverslip. The participant grew the in situ biofilms oncustom-made glass slabs fitted into a removable splint. The splintwas worn the entire time except during meals, and was immersedin 10 % sucrose three times a day. Unsterile stimulated saliva and4 % sucrose were used as flow medium, and the pH was analyzedusing the ratiometric dye C-SNARF-4. The results of this pilotstudy showed that after 30 min of static conditions, pH ranged from5.7 to 6.9, with differences both between fields of view (FOVs) andbetween biofilms, confirming the presence of microenvironments.After the onset of flow (5 mm/min), pH rose to neutral orslightly alkaline values in all three biofilms. However, differencesbetween FOVs were still present. In conclusion, incorporatingflow is important when studying pH developments inside biofilms.

AB - Metabolic processes inside dental biofilms are likely to be influencedby the velocity of the thin saliva film in contact with the biofilm.In addition, in situ-grown biofilms are likely to behavedifferently than in vitro biofilms. However, there is a lack of commerciallyavailable flow-cells able to incorporate in situ samplesand adjust the saliva film to a match the oral cavity. Therefore, theaim of this study was to design and 3D-print such a microfluidicflow-cell. As a proof of concept, we studied pH developmentsinside three 96 h biofilms under a stimulated saliva flow. Themicroscope compatible flow-cell consists of an incoming port, outcoming port and a central bottomless impression for the in situsample. Sealing the flow-cell to a coverslip forms a bottom to theflow-cell and creates a defined flow-space between the in situ sampleand the coverslip. The participant grew the in situ biofilms oncustom-made glass slabs fitted into a removable splint. The splintwas worn the entire time except during meals, and was immersedin 10 % sucrose three times a day. Unsterile stimulated saliva and4 % sucrose were used as flow medium, and the pH was analyzedusing the ratiometric dye C-SNARF-4. The results of this pilotstudy showed that after 30 min of static conditions, pH ranged from5.7 to 6.9, with differences both between fields of view (FOVs) andbetween biofilms, confirming the presence of microenvironments.After the onset of flow (5 mm/min), pH rose to neutral orslightly alkaline values in all three biofilms. However, differencesbetween FOVs were still present. In conclusion, incorporatingflow is important when studying pH developments inside biofilms.

M3 - Conference abstract in journal

JO - Caries Research

JF - Caries Research

SN - 0008-6568

Y2 - 9 July 2020 through 11 July 2020

ER -