IGEM:IMPERIAL/2008/Prototype/Drylab/Motility data collection

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m (Microscope Video Characteristics)
m (Microscope Video Characteristics)
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**The images are noisy
**The images are noisy
**Some cells seem to be undergoing division
**Some cells seem to be undergoing division
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**Some cells appear to be dead as they are not moving or drift

Revision as of 13:17, 5 September 2008

Home The Project B.subtilis Chassis Wet Lab Dry Lab Notebook


Contents

Our Approach

Material

We will be using the Zeiss Axiovert 200 inverted microscope with a fully motorised stage, controlled by Improvision Volocity acquisition software. This system offers a full incubation chamber with temperature and CO2 control, a large range of filter sets from UV to far-red and a highly sensitive 1300x1000 pixel camera for fast low-light imaging.

Video images are captured into memory by the system at a basal video frame rate of 16.3Hz. This can be further increased by performing binning.


Microscope Video Characteristics

  • From our experience on the microscope, we observe the following:
    • Bacteria are rod shaped.
    • Some cells swim out of the focus plane
    • The images are noisy
    • Some cells seem to be undergoing division
    • Some cells appear to be dead as they are not moving or drift


Video1

Video2

Video3

Manual Tracking Methodology

  • We intend to track for each cell:
    • The centroid
    • The part of the cell in the direction of movement
  • We attempt to reduce the bias in our manual tracking
    • By disabling the view path option
    • By displacing the cursor away from the tracked cell before adding a new point

Data Clustering


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