interLab

Objectives

While bacman is gearing up to protect people from arsenic , we decided to pay a visit to the gfp expressing bacteria. It s no doubt that measuring flourescence sounds exciting but even more appealing was the opportunity to be a part of a large community and share data.

So we prepared ourselves to answer the Intelab question : Can we reduce lab-to-lab variability in fluorescence measurements by normalizing to absolute cell count or colony-forming units (CFUs) instead of OD?

And then we started :
Day 1 : Transforming E coli DH5 alpha cells with the required biobr/icks.
Day 2 : Checking plates . Some biobr/icks didn’t give enough colonies.
Day 3 : Transforming bacteria with 1.5 uL of biobr/icks. Checking plates. We had beautiful and ample colonies this time.
Day 4 : Performing all calibr/ation protocols with ludox , silica beads , fluorescein.
Day 5 : Performed the cell measurements.
Day 6 : Repeated cell measurements.
Day 7 : Performing CFU protocol.
Day 8 : Counting colonies and updating excel files.

Results

Fluoroscence Raw Readings

Abs600 Raw Readings

Observations

• The test device 3 doesn’t show a great increase in fluorescence though its absorbance is increasing. Its expression level thus must be low.
• The test device 2 has shown difference florescence and absorbance for the two difference colonies. This could be indication of improper growth in one colony.
• The test device 1 has shown very low growth after 6 hrs. This could be due to slow growth of bacteria in set conditions and time.

Conclusion

At the end of interLab we learnt not only how difficult it might be to standardize data and yet how important this process is. We were glad to be a part of the interLab study and contribute in some way what was required in the effort to standardize data.

End result

Team IISER Kolkata is happy and satisfied with interLab.