Hello, can anyone advice me how can I determine Clostridium difficile growth using a spectrophotometer? Is there a specific dye which can be used (like the way resazurin can be used for aerobic bacteria)? I tried determining the bacterial growth in the different wells of 96 well plates using OD at 600 nm but the standard deviation for the wells that has seemingly consistent growth are very high.
It is a standard method to measure OD at 600 nm with regular time interval for the assessment of bacterial growth.
C. diff is a picky grower. OD600 with TSB works well, just make sure each well is getting the same lack of exposure to oxygen. If it's in a shaker, or not, look out for liquid forming a seal in the top of each well. This can cause a sealed aerobic well and slow growth compared to other wells.
On the other hand, it could be to endospore formation. Are you reactivating spores or ignoring them?
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