Another Project Update

I’m kinda stuck with my project. So, the situation is as follows: I have beamtime at Diamond Light Source in January, continuation report submission deadline in February and first year viva in March. Everything is good apart from both of my scaffolds not working. LOL :D In my project I have bioactive glass fibres and collagen gel on which I culture cells. The collagen gel doesn’t form properly and my supervisor will come to the lab with me this week to make sure my protocol is correct. If I am doing everything right it is most likely that we got faulty batch of collagen. Honestly, the collagen looked weird as it got all clumpy and very viscous. I called the supplier and they told me to heat it up in a water bath for 30 min to make it liquid again. To be honest its doesn’t sound right as we didn’t have to do any of that with the previous batch. Also, when I prepare collagen gel I rely on change of temperature to initiate gelation. So normally I would get it at 37, not make it liquid. Obviously, you need buffers, right pH etc. but temperature is a major step in my protocol. So, I will try to do it with my supervisor and then I will know what to do. This is not as big of a problem as the cotton wool like fibres. :D

With the fibres, its more complicated. Firstly, I run out of them so I need to spin more. All my previous batches I made at Harwell, but now I am trained to do it in Manchester. Next Wednesday I will spend a day trying to spin them with my BSc student. Anyway, the diameter of fibres that I had is too small so we won’t be able to see them during my beamtime. Furthermore, there are a ton of polymer drops which significantly obstruct the view. My cells also seem to die when I culture them on the fibres. I have tried to precondition them but it didn’t change anything. I also done it wrong, you are not supposed to use FBS supplemented media but whatever. :D At the moment I am doing a study where I compare old batch of fibres that had significantly larger diameter to new thin fibres. If the old batch gives us a better result it will mean that cells are killed as they pierce themselves as they try to engulf the fibres. Also, there is a small chance that I wasn’t washing them properly with PBS after sterilizing in ethanol and residual ethanol kills the cells. We will see, anyway I have to spin more fibres. So, the conclusion is that I have beamtime for which I don’t have any scaffold and my viva is approaching and none of my experiments worked. I can only explain what will be my future steps to make it work.