Team:Newcastle University/Conclusions

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To analyse the results of the wet lab transformations of the inserts into ''B. subtilis'', we used two methods: [[fluorescence microscopy]] and flow cytometry.
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To analyse the results of the wet lab transformations of the inserts into ''B. subtilis'', we used two methods: microscopy and flow cytometry.
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Microscopy work from the 08.09.08 showed a difference in the level of flourescence of the iGEMgfp fluorescent cells (higher in 10% subtilin-induced cells compared to 0% subtilin-induced cells). However, there was little difference in the ''number'' of cells that fluoresced between the two cultures.
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'''0% induction by subtilin''' (i.e in the absence of subtilin): mean flourescence = 7.70
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'''1% induction by subtilin:''' mean flourescence = 14.77
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'''10% induction by subtilin:''' the mean flourescence = 21.95
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These results show that the higher the concentraion of subtilin, the more GFP is expressed.
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There was no difference in the number of fluorecent cells ''or'' the level of flourescence between the 10% subtilin-induced and the 0% subtilin-induced iGEMcherry cells.
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Microscopy work done on 08.09.08 showed a possible difference in the brightness of the iGEMgfp fluorescent cells (brighter in 10% subtilin-induced cells than in 0% subtilin-induced cells). However, there was little difference in the numbers of cells that fluoresced between the two cultures. There was no difference in either the number of fluorecent cells or their brightness between the 10% subtilin-induced and the 0% subtilin-induced iGEMcherry cells.
 
===Flow cytometry===
===Flow cytometry===

Revision as of 13:55, 19 September 2008

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Newcastle University

GOLD MEDAL WINNER 2008

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Home >> Results

Results

Microscopy results of iGEMgfp upon 1% induction by subtilin
Microscopy results of iGEMcherry upon 1% induction by subtilin
Flow cytometry results for 1% induction by subtilin.
Flow cytometry results for 10% induction by subtilin.

To analyse the results of the wet lab transformations of the inserts into B. subtilis, we used two methods: microscopy and flow cytometry.

Microscopy work from the 08.09.08 showed a difference in the level of flourescence of the iGEMgfp fluorescent cells (higher in 10% subtilin-induced cells compared to 0% subtilin-induced cells). However, there was little difference in the number of cells that fluoresced between the two cultures.

0% induction by subtilin (i.e in the absence of subtilin): mean flourescence = 7.70 1% induction by subtilin: mean flourescence = 14.77 10% induction by subtilin: the mean flourescence = 21.95

These results show that the higher the concentraion of subtilin, the more GFP is expressed.

There was no difference in the number of fluorecent cells or the level of flourescence between the 10% subtilin-induced and the 0% subtilin-induced iGEMcherry cells.


Flow cytometry

Flow cytometry allows us to quantify our results and present them in graphical form. A sample of cells our engineered Bacillus subtilis cells were injected into the machine which hydro-dynamically focusses the fluid. Lasers are directed onto the stream of fluid, and each particle which passes through the light beam will cause the laser to scatter in a particular way. Fluorescent chemicals are excited to a higher energy state.

The detectors in the machine measure the scattering of light and any flourescence which occurs.