This is the homepage for 27820 - Ph.D course in Medical Biofilm Techniques.
Note: This page will be continuously updated and you should not expect all manuals to be here before a few days before the course starts.
Here you can find the general description of the course: DTU Course base
The course will start August 13th 2013 at 9.00 am in room 056 (ground floor) at DTU, Building 301. For directions, please follow this link.
Click here for the General time table in short, version 1.20 Subject to changes. (PRINTED)
- Laboratory rules for work in the P2 Lab Download and READ (PRINTED)
- Some files require a password to download. This should have been provided to you along with the link to this page. If for some reason you have not received the password, please e-mail Claus Sternberg.
- Manuals and other materials marked with "(PRINTED)" will be given in hard copy at course start
FACEBOOK: We have a Facebook group for present and past participants and instructors/speakers which you can join if you like. Go here: http://www.facebook.com/groups/120786086637/ and apply for membership. Note: The group is not terribly active, but who knows, perhaps someone will make it more interesting some time... :-)
Here we will keep you updated on changes of schedule, and other things you need to know (e.g. where to find result data files)
11-08-2013. Participant list uploaded.
14-08-2013. Participant List updated.
16-08-2013. Added motility assay protocol and speaker presentations.
18-08-2013. The title of the talk of Søren Molin will be: "Pseudomonas aeruginosa biofilms in CF airways – the role of adaptive mutations".
19-08-2013. The talks will start at 10.30. Supervisors (Janus and Claus) will be around from 9.00
Thank you to all the students of 27820 PhD course 2013. It was a great pleasure having you here!
Here we will put changes and errors we detect in the documentation. This document will be updated as the course is progressing (latest update date is shown.
Manuals for the excersises
- Biofilm System 1
- E.coli development: Download (updated Aug 7) (PRINTED)
- Biofilm System 2
- P. aeruginosa: FISH and Cell counting: Download (PRINTED)
- Biofilm System 3
- P. aeruginosa: Differentiation and colistin tolerance: Download (PRINTED)
- Biofilm System 4
- Monitoring gene expression in microbial biofilms: Download (PRINTED)
- Biofilm System 5
- Free exercise: Download
- Microbial Diagnostics
- Chip technlology and antibiotics screening (Exercise 6): Download (PRINTED)
- Presentation of results (Exercise 7)
- How to present on the last day: Download
Supplementary manuals and recipes
- Biofilm Media Download
- Biofilm general manual Download
- FISH Manual Download
- Embedding and FISH on flow chamber biofilms Download
- Adhesion Microtiter Tray Assay (Crystal Violet) Download
- Quorum Sensing assays (from course 27254) Download
- LIVE/DEAD kit manual Download
- Motility assay Download
- Cheat sheet for making Motility assay plates (Excel sheet) Download
- Diagnostic Excersise
- Short manual for analysis of AT Chip data Download
- Excel spreadsheet for AT Chip data Download
- Data Analysis Example Download
- Reading E-Test (Guide) Download
- Manuals for the BIOLOG machine
- OmniLog ID User Guide Download (See pages 42-44)
- OmniLog PM User Guide Download
- GEN III Database List and Characteristics Download
- MicroStation System Version 4.2 User Guide Download
Papers of relevance
Note: These papers are also included in the zipped package which you can download below.
- Heydorn, A., Nielsen, A.T., Hentzer, M., Sternberg, C., Givskov, M., Ersbøll, B.K., Molin, S. (2000): Quantification of biofilm structures by the novel computer program COMSTAT. Microbiol. 146(10): 2395-407 (The original Comstat1 paper)
- Heydorn, A., Ersbøll, B.K., Hentzer, M., Parsek, M.R., Givskov, M., and Molin, S. (2000): Experimental Reproducibility in Flow-Chamber Biofilms. Microbiol. 146(10): 2409-15 (The first paper using Comstat)
- Tolker-Nielsen, T., and Sternberg, C.(2011): Growing and Analyzing Biofilms in Flow Chambers. Curr. Prot. Microbiol. Unit 1B.2 (PRINTED)
(The papers are in a zip file on our secure server. Password protected)
- Supplementary papers (general) Download here
- Supplementary papers (Diagnostic exercise) Download here
- Motility paper by George O'Toole et al 1998 Download here
Result Data Files
Your own presentations!
- Download all the presentations from 2013 here
Biofilm System 1
Biofilm System 2
Biofilm System 3
Biofilm System 4
Biofilm System 5
- Check images on day one (Leica microscope)
- Day 2 (Zeiss microscope)
- Various exp 5 files (the file is called biofilm4.mdb.zip, funny enough)
List of Speakers and Notes
Powerpoint notes will be placed here as they become available
Speakers and teachers
- Claus Sternberg
- Janus Haagensen
- Lea Madsen
- Fatima Yousef
- Lars Jelsbak
- Søren Molin
- Anders Folkesson
- Claus Moser
- Lars Behrend
- Klaus Kirketerp-Møller
- Thomas Bjarnsholt
When all that has been done, get the comstat2 program file (Comstat2_.jar): here .
Note: When you download the comstat2 program file we imply that you accept the license conditions which are set out on www.comstat.dk, namely:
- That you will not distribute the program to others, but refer them to www.comstat.dk where they can obtain a copy for free
- That you will acknowledge the use of Comstat2 in any publication that will be written using Comstat2 generated data in any form.
Download the program file appropriate for your system:
- Imaris 7.6.4 Microsoft Windows XP/Vista/7 32 bit
- Imaris 7.6.4 Microsoft Windows XP/Vista/7 64 bit
- Imaris 7.6.4 Apple MaxOSX
- Demo license for the full Imaris Package - note: Right-click on the link and select "Save as..."
Install the program file. Save the license file somewhere on your computer and when you start Imaris, use the browse button in the license dialog box shown to point to the demo license file.
The demo license is valid until the end of August 2013. If you need testing after that, please contact Bitplane.
See also: Installation Instructions on bitplane.com
In order to enable Imaris to communicate with MATLAB, ImageJ and Java, you need to download and install these two helper programs:
Note: In this course it is unlikely that you will need the MATLAB/Java functionality of Imaris. But in case you want to check it out we include the links.
and the MATLAB Compiler Runtime (MCR). Please download the MCR version appropriate for your system:
You may also want to download SceneViewer which allows you to show Imaris stacks without a full license. You will, however, need to obtain a permanent license directly from Bitplane, Inc. The program will show how to get that when the demo license has expired.
- SceneViewer 7.6.4 Microsoft Windows 32 bit
- SceneViewer 7.6.4 Microsoft Windows 64 bit
- SceneViewer 7.6.4 Apple MacOSX
Brochures for Imaris:
Zeiss ZEN File Viewer
You can download the viewer program for native Zeiss confocal image stacks here
Leica LAF Viewer
You can download the viewer program for native Leica confocal image stacks here
Analysis of AT chip Images
A plugin for ImageJ have been developed to analyse the images from the AT Reader. The plugin evaluate the pattern and produce the corresponding "bar code". The plugin can be downloaded by clicking here (AT Chip Analyzer Plugin, and a manual is provided by clicking here (plugin manual).
In short: First install ImageJ (http://rsbweb.nih.gov/ij/). Then place the plugin in the plugin folder of ImageJ. And you are then able to run the plugin from ImageJ's plugin menu.
The flow cell assembly and flow system setup has been documented in JoVE (Journal of Visualized Experiments).
We have made this paper freely accessible without subscription:
- Weiss Nielsen, M., Sternberg, C., Molin, S., Regenberg, B. (2011), Pseudomonas aeruginosa and Saccharomyces cerevisiae Biofilm in Flow Cells. http://www.jove.com/details.php?id=2383 doi: 10.3791/2383. J Vis Exp. 47 (2011).