Team:SDU-Denmark/project-p

From 2010.igem.org

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To be able to observe an effect in the modified bacteria we had to expose the cultures to light, which was done through the microscope's integrated light source, which could switch between blue, green and ambient light. To further make sure that the blue light was at the correct wavelength the same optical filter that was used for the plate experiments was inserted between the lightsource and the microscopy slide. Since microscopy is impossible without light, we had to use red light as a replacement for darkness, the source of the red light was ambient light from the microscope's light source filtered through a red optical filter (about 580nm). This should not have any effect on the phototactic bacteria according to Jung et. al (insert reference!). The microscopic slides were prepared by adding 5 ul of bacterial culture in the center of a glass microscopy slide, dimensions 7,5cm x 1,5cm, at last a cover slip was used to cover the culture. This was then sealed with ordinary nail polish to eliminate any flow in the system, which could be mistaken for bacterial motility. The room where the experiment was carried out was kept as dark as possible, as to eliminate the chance of the phototactic bacteria being unwantedly exposed to light. The actual experiment was carried out in the following manner:<br><br>
To be able to observe an effect in the modified bacteria we had to expose the cultures to light, which was done through the microscope's integrated light source, which could switch between blue, green and ambient light. To further make sure that the blue light was at the correct wavelength the same optical filter that was used for the plate experiments was inserted between the lightsource and the microscopy slide. Since microscopy is impossible without light, we had to use red light as a replacement for darkness, the source of the red light was ambient light from the microscope's light source filtered through a red optical filter (about 580nm). This should not have any effect on the phototactic bacteria according to Jung et. al (insert reference!). The microscopic slides were prepared by adding 5 ul of bacterial culture in the center of a glass microscopy slide, dimensions 7,5cm x 1,5cm, at last a cover slip was used to cover the culture. This was then sealed with ordinary nail polish to eliminate any flow in the system, which could be mistaken for bacterial motility. The room where the experiment was carried out was kept as dark as possible, as to eliminate the chance of the phototactic bacteria being unwantedly exposed to light. The actual experiment was carried out in the following manner:<br><br>
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1. Expose bacteria to red light for 1 minute, while recording video.
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1. Expose bacteria to red light for 1 minute, while recording video.<br>
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2. Move to another spot on the slide, repeat 1. (repeated until 5 videos were recorded)
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2. Move to another spot on the slide, repeat 1. (repeated until 5 videos were recorded)<br>
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3. Expose bacteria to blue light for 1 minute, while recording video.
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3. Expose bacteria to blue light for 1 minute, while recording video.<br>
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4. Wait for 30 seconds, for the bacteria to reset themselves.
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4. Wait for 30 seconds, for the bacteria to reset themselves.<br>
5. Move the focus to another spot on the slide, repeat 3. (x5)<br><br>
5. Move the focus to another spot on the slide, repeat 3. (x5)<br><br>
This was done with all three strains of bacteria.<br>
This was done with all three strains of bacteria.<br>
An example of the results: <br><html>
An example of the results: <br><html>
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<object width="425" height="344"><param name="movie" value="http://www.youtube.com/v/HEKbYjyWNJc?hl=de&fs=1"></param><param name="allowFullScreen" value="true"></param><param name="allowscriptaccess" value="always"></param><embed src="http://www.youtube.com/v/HEKbYjyWNJc?hl=de&fs=1" type="application/x-shockwave-flash" allowscriptaccess="always" allowfullscreen="true" width="425" height="344"></embed></object></html>
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<object width="300"><param name="movie" value="http://www.youtube.com/v/HEKbYjyWNJc?hl=de&fs=1"></param><param name="allowFullScreen" value="true"></param><param name="allowscriptaccess" value="always"></param><embed src="http://www.youtube.com/v/HEKbYjyWNJc?hl=de&fs=1" type="application/x-shockwave-flash" allowscriptaccess="always" allowfullscreen="true" width="300"></embed></object></html>
==== K343004 ====
==== K343004 ====

Revision as of 01:46, 24 October 2010