what was your hardest experiment? – Disease Detectives Zone

Question: what was your hardest experiment?
- Keywords:
  difficult,
  experiment
Asked by anon-254469 on 14 May 2020.
- Emma Daniels answered on 14 May 2020:
  
  Hello Jenny, what a great question! I find all experiments difficult the first time I do them. Each experiment needs different equipment, or requires different knowledge, and I always find it hard and a bit scary to do something I haven’t done before. But, like most things, I find it gets easier the more I practice.
  
  I think the hardest experiments I do use a Schlenk Line. This is a piece of equipment you use when you’re working with chemicals that degrade in air or in contact with water. You attach your reaction flask to one of the lines and put it under vacuum so it removes all the air. You then refill your flask with an inert gas, such as argon or nitrogen, and then do your reaction. You have to make sure that your flask isn’t opened to air at any point, or it might ruin your reaction! I found it really difficult to use at first, and I still have to concentrate when using it now!
- Helena Fisk answered on 14 May 2020:
  
  Hi Jenny, Working with cells can be tricky as they are very demanding! Growing cells in the lab takes a lot of care and patience. We need to grow them in sterile conditions known as aseptic in special sterile laminar flow hoods (where even the air is sterile) to make sure the area is completely sterile (free from bacteria, viruses, fungi, DNA, general dirt, anything you can think of really) to make sure we do not introduce any infection or anything that can affect the growth or kill our cells. We then have to keep them at a certain temperature in an incubator with a certain level of oxygen which is dependant on what type of cells you’re trying to grow. We also have to make sure we feed them and change the liquid we grow them in that contains their nutrients (called media) to keep them happy. Also as they grow they get very crowded so we have to split them into new flasks to give them more room.
  Occasionally some users who are not so careful can bring something not sterile on to equipment or in to the incubator and it can cause mould or other nasty things to grow in other peoples’ cells and kill them! Other times you can do everything you think correctly but they just have a bit of a tantrum and don’t grow very well, or faster than usual and get overcrowded and stressed. this can be frustrating as some cells can take almost a month to grow for experiments!
  Cells are divas! They’re very fussy haha
- Sandra Greive answered on 14 May 2020:
  
  Wow, that’s tough question. I’ve done a lot of experiments that were difficult. As Emma says, they’re even harder the first few times as it takes practice to get good at doing new types experiments. Also sometimes we don’t quite know about the system will work and we have to try a few different things to figure out what works the best. The hardest experiments I’ve had to do were trying to make virus like particles for a Hepatitis C Virus vaccine. The aim was to make a particle that looked like the outside of the virus, but was not infectious. This was really hard to do because we needed to use mammalian cells to make the virus proteins and these are not as efficient at making tons of protein as bacteria, so the yield was very low. I had to try lots of different variations and combinations of the proteins to see what would assemble into particles. Each different test had to be analysed in a long and complex series of purifications to see if there were any virus like particles present. One run could take two weeks of really hard work before getting any kind of answer. I did manage to make large complexes that looked like they might have been virus-like particles, but then my funding ran out, and that was the end of the project. Turns out vaccines for hepatitis C virus (HCV) are really hard to make and not as cost-effective or preventative as reducing transmission by changing behaviour that minimised opportunities for blood contamination and transfer. Things like testing the donated blood supply for HCV infection, implementing protective barriers in health care, and educating IV drug users about not sharing needles etc has worked really well.
- Alex Holmes answered on 14 May 2020:
  
  Hi! I think the experiment that felt the hardest was one of the first ones I ever did as a masters student. I had to get some DNA inside of some E.coli, which is a pretty routine thing to do in the lab. However, it was my very first time and I didn’t want to mess up in front of my new lab mates! To do the experiment properly you have to keep everything sterile, so every bottle and tube needs to either be kept closed or only opened really close to a bunsen burner flame. To keep things close enough to the flame you need to be holding them – which is really tricky when you’re suddenly holding tubes in one hand and a pipette in the other and having to take lids off with the same hand you’re holding the tubes in and stay close to the flame but not burn yourself. It took a bit of time to get coordinated enough to do it quickly!
- Judith Sleeman answered on 14 May 2020:
  
  Everything is hard the first time, especially experiments with lots of different steps over several days, but some things stay a bit tricky. Micro-injecting cells with chemicals is probably the fiddliest thing I do. You have to get the chemical into a very fine glass tube, then jab each cell with the needle, while pushing compressed air through the tube. Jab too hard and the glass shatters, use too much air pressure and the cells blow up like balloons and explode!
  
  When I was doing my PhD, I used to run really giant gels loaded with lots of radioactivity. I’m short, so my arms were only just long enough to make the gels and the radioactivity used to make me really nervous, so that was physically hard. Most things do become much easier as you repeat them, but my arms never did get long enough to handle those gels easily!
- Laura Durrant answered on 14 May 2020:
  
  Hey Jenny!
  My trickiest experiments were probably the ones I carried out for my masters research project. I was testing a drug on human cells that I was growing in the lab. I had to add teeny-tiny amounts of drug solution (1ul to be precise) to up to 96 cell-containing wells at a time… plus, my samples were treated with a fluorescent tag, so I had to do this with the lab lights switched off! It was very finicky and tricky to do in minimal light… it required all of my concentration. Not to mention, sometimes my cells would just die on me for no apparent reason, which was nice. But I worked on my technique and my results started getting better! Sometimes it just takes practice.
- Alena Pance answered on 15 May 2020:
  
  It took me a long time to learn how to make red blood cells from stem cells. But that wasn’t enough, I needed to infect them with the malaria parasite to make my project work. To do that, I had to make parasites glow in different colours so that I can see them with a special machine called flow cytometer. But even when I got that to work, I still needed to tweak the flow cytometer to actually ‘see’ the parasites so that I can count the number of them that infected my made up red blood cells. And I couldn’t have done it without my colleagues who taught me things, gave me clues and suggestions and overall helped me to get it all to work and complete the project.
- Louisa Lee answered on 20 May 2020:
  
  I agree with the other scientists that say that it’s difficult the first time you do any experiment but specific types of experiments can be extra challenging. I used to do metabolomics experiments, which involved having to firstly create a condition for the cells to grow in, then quickly stop it and break open the cells while keeping them cold so that I could extract all the molecules for analysis. This was very fiddly, because the liquid I needed to collect the molecules in had alcohol in it, which meant it would evaporate very quickly!

Question: what was your hardest experiment?

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