Difference between revisions of "Part:BBa K412000:Experience"

Revision as of 03:27, 28 October 2010

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Applications of BBa_K412000

All images were taken with Olympus IX81 automated inverted microscope specially equipped for live cell imaging. All experiments were performed with DH5α E.coli cells. The filter sets we used are: 545/30x (excitation) and 620/60m (emission) filters for DsRed, 470/40x (excitation) and 525/50m (emission) for GFP. Data collection and processing was performed by the SlideBook software.
The data was taken using cells that were grown overnight in 2mL of LB broth that had 50ug/mL of the appropriate antibiotic at 37C and 220rpm. This was used in a 1:50 dilution. Each sample had 40uL from the broth grown overnight and 2mL of the LB broth with the appropriate antibiotic. These were allowed to grow until a predetermined OD. Then the contaminants were added and the cells were allowed to grow for another 2 hours, before the images were taken.
The images shown are the images used to get the data. From left to right the image is the negative control to a 10mM concentration of the respective contaminants. The graph shows the average of all the maximum intensities of the cells in the image vs the concentration of nitrate/nitrite added to the sample. The data shows that PyeaR+GFP responds better to nitrates than nitrites. It also shows that the cell is more sensitive to aluminum nitrate nonahydrate, and therefore starts dying at a lower concentration.

The images above are timelapse images. The images were taken every ten minutes after the 10mM potassium nitrate was added to the sample.

User Reviews

UNIQ27cd7c7a096d521a-partinfo-00000000-QINU UNIQ27cd7c7a096d521a-partinfo-00000001-QINU

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 ===Applications of BBa_K412000===
 [[Image:ver2.jpg]]
+*All images were taken with Olympus IX81 automated inverted microscope specially equipped for live cell imaging. All experiments were performed with DH5α E.coli cells. The filter sets we used are: 545/30x (excitation) and 620/60m (emission) filters for DsRed, 470/40x (excitation) and 525/50m (emission) for GFP. Data collection and processing was performed by the SlideBook software.
+*The data was taken using cells that were grown overnight in 2mL of LB broth that had 50ug/mL of the appropriate antibiotic at 37C and 220rpm. This was used in a 1:50 dilution. Each sample had 40uL from the broth grown overnight and 2mL of the LB broth with the appropriate antibiotic. These were allowed to grow until a predetermined OD. Then the contaminants were added and the cells were allowed to grow for another 2 hours, before the images were taken.
+*The images shown are the images used to get the data. From left to right the image is the negative control to a 10mM concentration of the respective contaminants. The graph shows the average of all the maximum intensities of the cells in the image vs the concentration of nitrate/nitrite added to the sample. The data shows that PyeaR+GFP responds better to nitrates than nitrites. It also shows that the cell is more sensitive to aluminum nitrate nonahydrate, and therefore starts dying at a lower concentration.
-We used our fluorescence microscope to take all of the images and to get the intensity data. Our results show that the part works as intended. When the concentration of nitrate or nitrite is increased the intensity of the fluorescence increases, up to the point where it is killing the E. coli. These results also show that PyeaR works better on nitrates, then nitrites, again as expected. The graph is of the mean intensity of all the cells in the picture at each concentration with the standard deviations shown.
+[[Image:tileview.jpg|800px]]
+The images above are timelapse images. The images were taken every ten minutes after the 10mM potassium nitrate was added to the sample.
 ===User Reviews===