I am growing B. subtilis in liquid culture, i see biofilm formation in 10-12 hours,. If i want to quantify cell growth, how should i do it using OD at 600nm? Since a lot of cells are a part of the biofilm and not freely available. Is there any manipulation i could do?
an easy way to Sessil count in the tube wall , it could be :
1. remove the micropipette population plantonica
2. Gently add buffer without touching the walls , but filling the tube.
3. Carefully remove all again .
biofilm population , should continue adhering to the walls , because its EPS should give you anchor , although extractions ( careful ) provided between the liquid pipetting .
Now again --to buffer with a known volume , and subjected to sonication tube with an internal tip to it. Check the parameters for release, not generate cell lysis.
Then remove an aliquot of the removal ,.
You can put in a tube for OD, remeber use other tube withoud bacterial only buffer for control.
Other way is staining and processed for counting by epifluorescence , as
BacLight staining LIVE / DEAD viability kit ( Molecular Probes Inc. , Eugene, Oreg . , USA ).
although you can use these two methods of counting in a complementary manner .
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