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Title: Design of a dynamic biofilm imaging cell for white-light interferometric microscopy

Abstract

In microbiology research there is a strong need for next generation imaging and sensing instrumentation that will enable minimally invasive and label-free investigation of soft, hydrated structures such as in bacterial biofilms. White light interferometry (WLI) can provide high resolution images of surface topology without the use of fluorescent labels but is not typically used to image biofilms because there is insufficient refractive index contrast to induce reflection from the biofilm’s interface. The soft structure and water-like bulk properties of hydrated biofilms make them difficult to characterize in situ, especially in a non-destructive manner. In this report, we build on our prior description of static biofilm imaging and describe the design of a dynamic imaging flow cell that enables monitoring the thickness and topology of live biofilms over time using a WLI microscope. The microfluidic system is specifically designed to create a reflective interface on the surface of biofilms while minimizing disruption of fragile structures. The imaging cell was also designed to accommodate limitations imposed by the depth of focus of the microscope’s objective lens. Example images of live biofilm samples are shown in order to illustrate the ability of the flow cell and WLI instrument to 1) support bacterialmore » growth and biofilm development, 2) image biofilm structure that reflects growth in flow conditions, and 3) monitor biofilm development over time non-destructively. In future work, the apparatus described here will enable surface metrology measurements (roughness, surface area, etc.) of biofilms and may be used to observe changes in biofilm structure in response to changes in environmental conditions (e.g., flow velocity, availability of nutrients, and presence of biocides). Furthermore, this development will open new opportunities for the use of WLI in bioimaging.« less

Authors:
 [1];  [1];  [1];  [1]
  1. Pacific Northwest National Lab. (PNNL), Richland, WA (United States)
Publication Date:
Research Org.:
Pacific Northwest National Lab. (PNNL), Richland, WA (United States)
Sponsoring Org.:
USDOE
OSTI Identifier:
1358486
Report Number(s):
PNNL-SA-123163
Journal ID: ISSN 0091-3286
Grant/Contract Number:
AC05-76RL01830
Resource Type:
Journal Article: Accepted Manuscript
Journal Name:
Optical Engineering
Additional Journal Information:
Journal Volume: 56; Journal Issue: 11; Journal ID: ISSN 0091-3286
Publisher:
SPIE
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; 47 OTHER INSTRUMENTATION; white light interferometry; biofilm; bacteria; non-destructive; imaging; flow cell; roughness

Citation Formats

Larimer, Curtis, Brann, Michelle, Suter, Jonathan D., and Addleman, Raymond Shane. Design of a dynamic biofilm imaging cell for white-light interferometric microscopy. United States: N. p., 2017. Web. doi:10.1117/1.OE.56.11.111708.
Larimer, Curtis, Brann, Michelle, Suter, Jonathan D., & Addleman, Raymond Shane. Design of a dynamic biofilm imaging cell for white-light interferometric microscopy. United States. doi:10.1117/1.OE.56.11.111708.
Larimer, Curtis, Brann, Michelle, Suter, Jonathan D., and Addleman, Raymond Shane. Wed . "Design of a dynamic biofilm imaging cell for white-light interferometric microscopy". United States. doi:10.1117/1.OE.56.11.111708. https://www.osti.gov/servlets/purl/1358486.
@article{osti_1358486,
title = {Design of a dynamic biofilm imaging cell for white-light interferometric microscopy},
author = {Larimer, Curtis and Brann, Michelle and Suter, Jonathan D. and Addleman, Raymond Shane},
abstractNote = {In microbiology research there is a strong need for next generation imaging and sensing instrumentation that will enable minimally invasive and label-free investigation of soft, hydrated structures such as in bacterial biofilms. White light interferometry (WLI) can provide high resolution images of surface topology without the use of fluorescent labels but is not typically used to image biofilms because there is insufficient refractive index contrast to induce reflection from the biofilm’s interface. The soft structure and water-like bulk properties of hydrated biofilms make them difficult to characterize in situ, especially in a non-destructive manner. In this report, we build on our prior description of static biofilm imaging and describe the design of a dynamic imaging flow cell that enables monitoring the thickness and topology of live biofilms over time using a WLI microscope. The microfluidic system is specifically designed to create a reflective interface on the surface of biofilms while minimizing disruption of fragile structures. The imaging cell was also designed to accommodate limitations imposed by the depth of focus of the microscope’s objective lens. Example images of live biofilm samples are shown in order to illustrate the ability of the flow cell and WLI instrument to 1) support bacterial growth and biofilm development, 2) image biofilm structure that reflects growth in flow conditions, and 3) monitor biofilm development over time non-destructively. In future work, the apparatus described here will enable surface metrology measurements (roughness, surface area, etc.) of biofilms and may be used to observe changes in biofilm structure in response to changes in environmental conditions (e.g., flow velocity, availability of nutrients, and presence of biocides). Furthermore, this development will open new opportunities for the use of WLI in bioimaging.},
doi = {10.1117/1.OE.56.11.111708},
journal = {Optical Engineering},
number = 11,
volume = 56,
place = {United States},
year = {Wed May 10 00:00:00 EDT 2017},
month = {Wed May 10 00:00:00 EDT 2017}
}

Journal Article:
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  • Both bacterial adhesion to surfaces and the subsequent formation of microbial biofilms have serious implications for a number of industrial applications in aqueous environments. The development of multicellular biofilms on submerged man-made structures such as drilling platforms and ship hulls is the first obvious stage in the fouling of these structures, which can progress to the point where costly cleaning or replacement is needed. On heat exchangers, even thin microbial films cause a serious decrease in efficiency of heat transfer. The polymers involved in the adhesion of Pseudomonas fluorescens H2S to solid surfaces were investigated to determine whether differences betweenmore » cell surface adhesives and biofilm matrix polymers could be detected. Two optical techniques, i.e., interference reflection microscopy (IRM) and light section microscopy (LSM), were used to compare the responses of the two types of polymer to treatment with electrolytes, dimethyl sulfoxide (DMSO), and Tween 20. Results indicate that both polymers bear acidic groups and thus act electrostatically with cations and are able to enter into hydrophobic interactions. Differences in their response to DMSO could be due to the close proximity of the glass surface or to differences in structure of the two polymers.« less
  • Bacterial biofilms are complex, three-dimensional, communities that are found nearly everywhere in nature1 and are being recognized as the cause of treatment-resistant infections1 2. Advanced methods are required to characterize their collective and spatial patterns of metabolism however most techniques are invasive or destructive. Here we describe the use of a combined confocal laser scanning microscopy (CLSM) and nuclear magnetic resonance (NMR) microscopy system to monitor structure, mass transport, and metabolism in active biofilms. Non-invasive NMR methods provide macroscopic structure along with spatially-resolved metabolite profiles and diffusion measurements. CLSM enables monitoring of cells by fluorescent protein reporters to investigate biofilmmore » structure and gene expression concurrently. A planar sample chamber design facilitates depth-resolved measurements on 140 nL sample volumes under laminar flow conditions. The techniques and approaches described here are applicable to environmental and medically relevant microbial communities, thus providing key metabolic information for promoting beneficial biofilms and treating associated diseases.« less
  • Chemical imaging of single cells is important in capturing biological dynamics. Single cell correlative imaging is realized between structured illumination microscopy (SIM) and time-of-flight secondary ion mass spectrometry (ToF-SIMS) using System for Analysis at the Liquid Vacuum Interface (SALVI), a multimodal microreactor. SIM characterized cells and guided subsequent ToF-SIMS analysis. Dynamic ToF-SIMS provided time- and space-resolved cell molecular mapping. Lipid fragments were identified in the hydrated cell membrane. Principal component analysis was used to elucidate chemical component differences among mouse lung cells that uptake zinc oxide nanoparticles. Our results provided submicron chemical spatial mapping for investigations of cell dynamics atmore » the molecular level.« less
  • A simple interferometric technique using both a white-light and a monochromatic-light source can be used to measure the wall thickness of hollow glass microspheres to better than 0.05 $mu$m. The entire procedure requires only a few minutes per sphere and can easily be made to take into account the effects of any gas with which the ball is filled. (AIP)
  • No abstract prepared.