Team:Groningen/Template/MODULE/Notebook/bandage/week15

The freeze dried cells made last week were used to prepare several gels for microscopy. From literature we found out that the pore size of a polyacrylamide gel of 2.5 % is approximately 200 nm and the pore size of a 10,5% gel is approximately 20 nm1

 

Also we found that the pore size of a 1.5% low melting point (LMP)agarose corresponds to a pore size of around 150 nm² as can be seen in figure 6. A L. lactis cell varies between sizes of 500nm up to 1500nm³

 

 

Figure 1: CAL1 spotted with B. subtilis 168, ATCC6633 grown in LB and ATCC6633 grown in SMM.

 

 

For these reasons we attempted to grow the L. lactis cells in a 2.5% polyacrylamide gel, a 5% acrylamide gel, a 1.5% LMP agarose gel and on the surface of a 5% polyacrylamide gel. Half an hour in advance of making the gel the cells were induced with nisin to start the GFP production. In this way we could find out whether the cells were still viable after polymerization of the gel they were poured in. The gels were observed and followed using a widefield (confocal) microscope. So far we can prove the cells are able to grow on top of a 1.5% LMP agarose gel as well as inside a 1.5% LMP agarose gel, results are shown in figure….

test 3