Chitozen

The 1st functionalized microscope coverslip to image live bacterial cells & study their growth and behavior. 

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Chitozen is a chitosan-coated coverslip compatible with 6-channel sticky slides.

What is it intended for?   Use it if you want to image bacteria both still and alive under the microscope. Or if you want to perform long-term imaging of bacteria. Or if you want to change the growth condition (e.g. antibiotics, chemicals, inhibitors) during the experiment and directly observe, in real-time, the bacteria new comportment under the microscope.

 For which bacteria?  E. coli, Vibrio cholerae, Myxococcus xanthus, Mycobacterium smegmatis, Bacillus subtilis, Pseudomonas aeruginosa and Pseudomonas fluorescens. More bacteria being currently tested by our users. Discover their feedback of use.

 Protocol, features and images

 Product information: Kit description, Lifetime, Technical and Safety datasheets, FAQ. 

Designed by  Tâm Mignot, Olivier Theodoly, Amandine Desorme, Guillaume Sudre and Laurent David.  Published in mBio
Discover the story of their tech transfer journey  

 See below 2 videos with E. coli.

How to use Chitozen


 
 
 
 

E.coli & Chitozen


 
  
 
 

E. coli monolayers on Chitosan
© 2019 Tréguier et al. This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.

Visualization of Pal mCherry at septum in E. coli (W3110 Pal mCherry), by 3D SIM microscopy, in M9 medium and using Chitozen.
Credit : Amandine Desorme - LCB - CNRS - 2021

Feedback from the community

Chitozen has been successfully tested by our Test Program users on the following bacteria: 
 E. coli
 Mycobacterium smegmatis
 Bacillus subtilis
 Pseudomonas fluorescens
 Salmonella.

NB: These results supplement those obtained by Tâm Mignot and al. with:
 E. coli
 Vibrio cholerae
 Myxococcus xanthus
 Pseudomonas aeruginosa.

Tracking of DNA repair proteins in live cells of Escherichia coli
By Maxence Vincent, University of Oxford, Jan 2022

"Cells expressing HaloTagged DNA repair proteins were loaded onto the chitosan-coated coverslip and imaged with TIRF microscopy. The Chitozen technology enables tracking single-molecules while changing environmental conditions (e.g: M9 or M9 + mutagen). Computation of the diffusion coefficients provides insights into how proteins change their mobility upon DNA damage".



 
  
Credit of all images and the video: Maxence Vincent - University of Oxford – 2022
Odoo • A picture with a caption


Time-lapse images of E. coli growing: Localization of F-plasmid
By Jerome Rech, LMGM-CBI-CNRS, Toulouse, 2022

Cells are observed in phase contrast (top), in the blue and yellow channels for fluorescence microcopy to observe phase contrast, nucleoid (DAPI) or F-plasmid (mVenus tag), respectively. 

Overlays of phase contrast and blue or yellow channel are shown on middle or bottom panels, respectively.

Cells were grown at 30°C in supplemented M9 minimal media containing Cystein and DAPI under continuous flow. Scale bar (2 µm). 

Odoo • Image and Text

Features

Full compatibility with most of your conditions of experiments

  Size: the coverslip dimension (25x75 mm) is compatible with the most common available sticky slides and microscope stages.

Compatible with advanced microfluidic techniques, nanolithography

6 independent channels


Either perform up to 6 experiments at the same time or use 1 channel one day, and the others later

Ready and fast

 Assemble it within 2 minutes
.
 Use it the same day it is prepared

Long lasting

.
 A bench-stable surface coated with chitosan, the most efficient way to immobilize your bacteria on a microscope coverslip

   Storable for 2 months once assembled

 

Gallery

The Chitozen video protocol  

 
 
Chitozen: how to use it, a video protocol from assembling the elements to imaging your favorite bacteria.

Simplified protocol





The material you will need:
Hardware

µ-Slide Microscopy Rack
Eppendorf® Centrifuge 5430 & Rotor 5430R A-2-MTP
Clamp and adapter for sticky slides 

Consumables (included in the Chitozen kit)

Bottomless 6 channels sticky slides (sterilized and welded in a gas-permeable packaging)

Reagents

LB medium
LB ½ medium 
Milli-Q® water

Cells

E. coli - Mycobacterium smegmatis - Bacillus subtilis - Pseudomonas fluorescens - Vibrio cholerae -  Myxococcus xanthus - Pseudomonas aeruginosa.

DOWNLOAD THE PROTOCOL

Product Information

5-Chitozen Standard kit
5 standard (25 x 75 mm) chitosan-coated coverslips with 5 sticky slides

1-Chitozen Starter kit
1 standard (25 x 75 mm) chitosan-coated coverslip with 1 sticky slide

Each Chitozen coverslip allows up to 6 assays using bottomless 6-channel sticky slides

Lifetime: up to 12-month storage at room temperature, shielded from direct sunlight.

Chitozen Technical Datasheet.
 Chitozen Safety Datasheet. 

Download the Chitozen FAQ.

Going further: perfuse the system with flow

Odoo • A picture with a caption
Chitozen coverslips allow bacterial cells to be immobilized even under a flow.

Chitozen and us

" We wanted to understand how our bacteria, Myxococcus xanthus, moves on a surface. So we tried to design a smart microscopy system to observe it. But there was a problem: the Myxococcus cells were not adhering to our glass slides. When we read a paper on how people use chitosan to get their bacteria better move on plates, we got an idea: maybe we could coat our glass slides with some chitosan? We tested it. And it worked! We then implemented our prototype with a microfluidics system to test the real time response of our bacteria to antibiotics. Now we’re thrilled to share it with the community of research! We hope that people working on bacteria will appreciate to use Chitozen because it is a very easy system to run that combines microfluidics and high-end microscopy. We are looking forward to getting feedback of use: it will mean that we have developed a device useful to the community and that is always a deep satisfaction! "
T âm Mignot, Olivier Théodoly, Amandine Desorme, Guillaume Sudre, Laurent David

 Contact  the developers about their technology
Odoo • Image and Text

Odoo • Image and Text Esther Graudens
New projects at Idylle
" We really loved the way all the team, whether in Marseille or in Lyon, was deeply concerned by the tiniest detail and willing to prepare the best ever product for the community. So if you want to know more about how we co-designed and produced Chitozen starting with chitosan, then catch an eye on   THE STORY OF CHITOZEN .
And if you are interested in seeing what you can do with chitosan-coated coverslips, you can have a look at this study published by the team of developers in 2019. They've managed to use chitosan-coated coverslips to promote the growth of cells without any deleterious effect on their physiology, allowing them to measure the antibiotic susceptibility of a diversity of clinical strains with an excellent accuracy in a very short period of time. Read all about it here: mBio , 2019"

Contact us about your challenges to image your bacteria
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