Macroscopic imaging of luciferase cells

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jamesonii
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Macroscopic imaging of luciferase cells

Post by jamesonii » Thu May 15, 2008 11:10 pm

Hi, I was wondering if anyone has any experience in macroscopic imaging (using a digital camera) of luciferase cells / any light emitting cell lines. Currently, I am using the Shh-light II cells from ATCC which is suppose to produce light emitting enzymes upon exposure to hedgehog proteins. These cells can then be collected and quantified using a luminometer. However, what we are trying to do here is to obtain a spatial map of the cell growth in a macroscopic level (to visualize the entire culture flask / bioreactor in a single image). I have tried to expose these cells under 20min exposure time and still do not achieve images at all. I have a feeling that since the cells are in the microscopic level and so are the enzymes, it will be difficult to visualize them at the macroscopic level. My supervisor tends to disagree as he thinks it should be like the firefly phenomenon, we shoudl be able to see this light with the naked eye. Call me crazy or him crazy? I need some advice. Thanks.

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canalon
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Post by canalon » Fri May 16, 2008 2:40 pm

I have limited experience but I once tried to work with a similar system to follow bacterial transformations. My 2 cents would be:
-Yes light emitted by bacteria can be seen, even by the eye (although after spending a while in the dark) so you should be able to capture it with a camera
- But IIRC the light emission is O2 dependant, and that can make things a little tricky, because sometimes ther is just not enough oxygen around your cells to create a detectable signal.
Patrick

Science has proof without any certainty. Creationists have certainty without
any proof. (Ashley Montague)

tittu
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Post by tittu » Sun May 25, 2008 2:46 am

try a 1 hr exposure and it must be totally dark all around when you expose, if not, longer :)

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