Bad news and a mistake in important experiment

So, last week was kind of important, I planned to prepare an experiment to determine cell seeding density for optimum vascularisation efficiency. It was quite straightforward, so I just had to prepare 6 samples each with different number of cells. The important part was that I do co-culture of two cell types and I use a ratio of 1:4. I started the experiment very confident and everything was going very smoothly. I had enough cells, the gel looked good and I always get a bit anxious when doing a cell count. So, I was sitting there and calculating the volumes of cell suspension that I need to add to each sample to obtain a required number of cells. A friend of mine was also working in a lab and he helped me out, by telling me how to do all those calculations much faster. I did and I think I got over excited that I can do them so fast, so I stopped concentrating. I completed the experiment and only when I got to my office to fill out my lab book I realized that I used 1:1 cell ratio. Which is kind of bad especially considering that one of my cell types is generally not very happy and will die off slowly. (Same in the literature) I checked literature straight away and I found that most people use 1:1 ratio so I hoped for the best. The reason why this is so bad is that I will be away next week, teaching so I won’t be able to do any cell work until mid-March and I was really hoping to get some good data, finally showing that somethings does work.

Another bad piece of news that I got last week was that my beamtime – time at Diamond Light Source to scan the samples, was moved. It initially was supposed to be in January, but no one was ready so we agreed that it will be in April and June. This or last week don’t exactly remember, I got a call from my co-supervisor notifying me that it was scheduled for 13 of March. I was in shock as I am away from 6th until 10th of March teaching and then I won’t even have any samples. My colleague with whom we wrote a proposal broke his elbow so he is not allowed to enter the labs at all, which is not great. So, I think they gave us 2 days in March and 2 days in April. I won’t have any samples for March I will try my best to prepare them for April, but I need to figure out cell density, if I don’t get blood vessel then there is no point. Also, the seeding density experiment was important as my co-supervisor asked me to stain my scaffolds and use them for teaching next week, if there is no organisation of cells I really doubt that it will be an interesting sample to teach. I don’t know, I am tired plus the braces are driving me mad. There is no pain anymore but my lips, OMG they are falling apart I have tried everything, they are chapped and hurt really bad. Lastly, I need to prepare for my viva but I fall asleep every time I start to read a paper so my progress isn’t great.