Microbiology & Biofilms

Long term non-invasive live cell imaging of microorganisms

Fungi: Trichoderma reesei spore germination

In this footage, which was obtained during a demonstration organized by Dr. Matthias Steiger at the Technical University in Vienna, we observe the germination process in T. reesei at 30 °C. One image was acquired every 5 min for 2 h 45 min; a time frame sufficient to observe the initial formation of the germ tube. The high resolution of the images permit visualization of the growing membrane, internal structures and the formation of the wall, which occurs after 2 h 20 min..

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Analysis of 3D colonies in bacteria

This image shows E.coli bacteria embedded in alginate beads (generated by Encapsulation Unit – Var J30, 40-70 microns diameter). The beads were mounted on a slide into Minimum Media (MM) diluted 1:4 with distilled water (plus 1% v/v Fumarate 100mM). The time-lapse imaging experiment was conducted using the 3D Cell Explorer with a standard top-stage incubator set to 37°C and 90% humidity for 3 hours, capturing images every 10 minutes.

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Analysis of 3D colonies in bacteria

This image shows E.coli bacteria embedded in alginate beads (generated by Encapsulation Unit – Var J30, 40-70 microns diameter). The beads were mounted on a slide into Minimum Media (MM) diluted 1:4 with distilled water (plus 1% v/v Fumarate 100mM). The time-lapse imaging experiment was conducted using the 3D Cell Explorer with a standard top-stage incubator set to 37°C and 90% humidity for 3 hours, capturing images every 10 minutes.

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Bacteria in tissues and cell samples

Rehydrated paraffin embedded bee gut sections (5 µm) imaged with the 3D Cell Explorer. The panoramic view of the whole tissue section (more than 400 micrometers) was obtained by stitching multiple acquisitions using a free online software. Like human intestine, bee gut is colonized by bacteria population. Most of the gut microbial community resides in the ileum.

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Macrophages phagocytosing E.coli

Phagocytosis Assay Kits by PromoCell were used to test the viability and cellular functioning of the macrophages (video 2 & 3). E.coli particles, visible as small ellipsoid particles, are trapped by the cells, transported and lysed. This system can be used to provide a robust screening system for activators and/or inhibitors of phagocytosis and Toll-like Receptor (TLR) ligands.

Read our detailed blogpost here.

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Macrophages phagocytosing E.coli

Phagocytosis Assay Kits by PromoCell were used to test the viability and cellular functioning of the macrophages (video 2 & 3). E.coli particles, visible as small ellipsoid particles, are trapped by the cells, transported and lysed. This system can be used to provide a robust screening system for activators and/or inhibitors of phagocytosis and Toll-like Receptor (TLR) ligands.

Read our detailed blogpost here.

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Scientific Publications

The 3D Cell Explorer allows for:

  • Label-free 3D infection process monitoring
  • 3D localization of bacteria inside living cells
  • Detection of mammalian cell infections
  • Volume measurements of intra-cellular parasites
  • Visualization of microorganism internalization process

Scientific Publication in Biofilm formation:

Impact of the Food Additive Titanium Dioxide (E171) on Gut Microbiota-Host Interaction

Authors: Gabriela Pinget, Jian Tan, Bartlomiej Janac, Nadeem O. Kaakoush, Alexandra Sophie Angelatos, John O’Sullivan6, Yen Chin Koay, Frederic Sierro, Joel Davis, Shiva Kamini Divakarla, Dipesh Khanal, Robert J. Moore, Dragana Stanley, Wojciech Chrzanowski and Laurence Macia

Published in frontiers in Nutrition

The University of Sydney & University of New South Wales have recently published their work on the “Impact of the Food Additive Titanium Dioxide (E171) on Gut Microbiota-Host Interaction”.

They have investigated the impact that Titanium Dioxide (E171) – mostly found in processed foods such as mayonnaise or candies – has on gut microbiota of mice. The study shows that the food additive E171 can induce gut microbiome changes associated with the development of diseases, including colorectal cancer and inflammatory bowel disease.

Visualizing bacteria and biofilms has been complicated with traditional microscopy due to low-contrast and high transparency of these samples. Thanks to the 3D Cell Explorer it has been possible to visualize E. coli and E. faecalis to reveal the clustering effect of TiO2 on both E. coli and E. faecalis in vitro in this study. Please see Figure 1 for reference.

TiO2 triggers biofilm formation by commensal bacteria. (A,B) The clustering effect of TiO2 on (A) E. coli and (B) E. faecalis in vitro was visualized by Nanolive imaging in the presence of 0, 0.5, 1, or 50 μg/ml TiO2 after 24 h incubation. Black represents the refractive index of TiO2 and green represents bacteria. From Gabriela Pinget et al., Frontiers in Nutrition., 2019

More Scientific Publications

Scientific publication “S-Nitrosothiol Plasma-Modified Surfaces for the Prevention of Bacterial Biofilm Formation“:

Authors: Zahra Sadrearhami, Rashin Namivandi-Zangeneh, Emily Price, Marta Krasowska, Sameer A Al-Bataineh, Jason Whittle, Edgar H. H. Wong, Anton Blencowe, Cyrille Boye

Published in ACS Publications. Find the publication here: Download.

Scientific publication covering the topic “An alternative method to determine the integrity of cells“:

Diagnosing water treatment critical control points for cyanobacterial removal: Exploring benefits of combined microscopy, next-generation sequencing, and cell integrity methods

Authors: Arash Zamyadi, Caitlin Romanis, Toby Mills, Brett Neilan, Florence Choo, Lucila A., Coral, Deb Gale, Gayle Newcombe, Nick Crosbie, Richard Stuetz, Rita K. Henderson

Published in Science Direct. Find the publication here: Download.

Scientific Publication in Biofilm dispersion:

Nitric Oxide-Loaded Antimicrobial Polymer for the Synergistic Eradication of Bacterial Biofilm

Authors: Rashin Namivandi-Zangeneh, Zahra Sadrearhami, Ali Bagheri, Maeva Sauvage-Nguyen, Kitty Ka Kit Ho,Naresh Kumar, Edgar H. H. Wong, and Cyrille Boyer

Published in: ASC Macro Letter.Find the publication here: Download.

Scientific Publication in Bacteria killing assessment & Biofilm dispersion assessment:

Enhancing the antimicrobial and antibiofilm effectiveness of silver nanoparticles prepared by green synthesis

Authors: Susan Oliver,  Harsha Wagh, Yuanli Liang, Shuang Yang  and  Cyrille Boyer

Published in:Journal of Materials Chemistry B. Find the publication here: Download.

Scientific Publication in Bacteria killing: 

The Effects of Polymer Topology and Chain Length on the Antimicrobial Activity and Hemocompatibility of Amphiphilic Ternary Copolymers

Authors: Rashin Namivandi Zangeneh, Rebecca J. Kwan, Thuy-Khanh Nguyen, Jonathan Yeow, Frances L. Byrne, Stefan H. Oehlers, Edgar Wong and Cyrille Boyer 

Published in: Polymer Chemistry Journal. Find the publication here: Download.

Video library

Bacteria growing in alginate beads

Pap-smear analysis

E.coli being engulfed through phagocytosis

Read our blogpost here.

 

Fungi: Aspergillus niger spore germination

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