Kirsty the Scientist

Today is my last day working in the lab and it feels very bittersweet!   Over the past 10 weeks, I've had the best time.  A huge part of that was down to working in the most amazing group of people. Not only showing me the ropes of lab work, but welcoming me into their team with open arms. I also had the opportunity to learn from a fantastic supervisor - someone who really knows what they are doing and guides you under their wing whilst allowing you to explore and develop your own ideas.  So, although I'm really looking forward to the next stage - getting my dissertation written up and tying all my story together - I'm going to really miss working in the lab every day. This experience has completely changed my perspective about lab life. I'd never realised just how collaborative, supportive and friendly the environment can be (if you work with a good bunch!). And I enjoyed lab work more than I thought too - I loved problem solving at the lab bench, considering differ...

"You're not a real scientist until you've spent 3 weeks working hard on something and at the end of it all you might as well have been at home in your pyjamas the whole time!"  Unfortunately this week has ended up being a bit of a write off (which is incredibly frustrating in such a short 10 week project!). My Western blots (the experiment used to detect the presence of specific proteins) have all not been working and we've been really stuck as to exactly why. But maybe I'll just have to except sometimes science doesn't work when we want it to! 

As I was talking about earlier this week, I've often been running more than one experiment at the same time during a week. But today, the differences between the two experiments seemed quite striking!  The two experiments are to look for two different indicators to suggest that my protein of interest is involved in making my cell type increase during cell stress/inflammation:  1. Look for the protein itself. This involves separating the proteins using electrophoresis (pulling the proteins through a polyacrylamide gel using an electric current to draw the proteins through the tiny pores of the bell - the smaller proteins get to the bottom of the gel quicker as they can wiggle through the holes faster. Then using a method called Wet Transfer, which pulls the protein out of the gel and onto a PVDF membrane - a technique that uses litres of buffer.  2. Looking for the genes that are expressed - suggesting that the cell is producing the proteins and using the part of the...

Forget everything I've said about being busy so far - today was the most hectic and experiment-filled day I've had yet!  There was a lot to get done today - finishing off longer experiments from last week, plating out cells ready for this week's experiments, data analysis of a couple of different experiments - all on top of a Masters student, who is still shadowing me, following my every move and asking questions about each step. All good multi-tasking practice, and reminding me why each step is important, but I had to be on the ball at all times! 

Normally, I only post new blogs on weekdays, but I've treated you all this week to a bonus weekend post to answer a question that I've kept quite under wraps throughout this blog - what is this project I'm working on actually about?! via GIPHY Well, for a start, there is a reason I have been a little bit elusive on this topic. Technically, the work I do belongs to the university, not me. Therefore officially, the information I gather is not my property to share with the public and the world - and this blog is open for anyone and everyone to read. And as I've said before, my subject area - as you'd expect - is quite niche, meaning this blog could be easily found with a quick google search if I was specific! Although I love how science is such a collaborative subject and sharing research can open your eyes to new angles and opinions on topics - it's not worth the hassle and the sabotage of my degree just to include the specifics of my project on the blog! ...

Today felt like a really productive day, despite all the lab work actually being very repetitive!  Got all the figures from the previous results looking really good today and sent them off to my supervisor to be looked over. And stimulate the cells at multiple timepoints across today, and made up the samples, ready for this week's experiments!  Busy, busy, busy! Definitely, eat, sleep, lab at the moment!

Similar to yesterday, (and most days nowadays!) I was working on two goals:  1. Prep more cells for use for a new experiment for this week.  2. Detect the secondary antibody signals using the Western blots using two slightly different methods (which I can compare to see which is better). Li-Cor detects the signals using fluorescence using a shiny new machine the institute has; whereas ECL is the standard method which uses light.  We also had our first full group meeting today, where the research team gets together and we can discuss the results of everyone's experiments - which will now become a bit more of a regular occurrence! 

Still processing the information gathered from last week's experiments (results collected on Friday). Reprobing the Western blots with more antibody to compare the two proteins we're looking for.  Also learnt how to use an 'alternative Excel' today - called GraphPad Prism, which is a bit better than Excel for producing the lovely good-looking diagrams and graphs ready for publication and to pop in my dissertation.  All in all, these first results make a good start to the project, and has given a few more clues as to the direction this project is going to take over the next couple of weeks. 

This evening I came home and almost instantly fell asleep! It's been a busy week, and a lot of information to take in each day and new things to learn! And today's results were a little anti-climatic, with some contamination in one of the lanes of the western blot, and the PCR results still a bunch of numbers that I'll be compiling into a graph over the weekend before they make much sense! 

Another busy day, preparing the plates of cells to be tested in 2 different experiments - one to search for the protein itself and one looking for the DNA that makes the protein.  On the cusp of finding out the results of a week's worth of work, I was reminded of something I listened to in a podcast today. It was a quote from Richard Feynman, the renowned physicist: "The scientist has a lots of experience with ignorance, doubt and uncertainty., and this experience is of very great importance. When a scientist doesn't know the answer to a problem they are ignorant, when they have a hunch as to what a result is they are uncertain, and when they are pretty darn sure of the result they are in doubt. Science is a satisfactory philosophy where doubt is not to be feared but welcomed and discussed." "To paraphrase - s cience is the enemy of the certain and being shown to be wrong is an invaluable part of learning about nature." And that's something I will k...

Today was the busiest lab day I've had so far! We needed to be back and forth between the tissue culture lab and our normal lab/the office at regular intervals throughout the day to continue the timecourse. I'm stimulating the cells at lots of different times to find out at which point the protein I'm looking for is produced. 

After this whole debacle of the lab relocation, the hunt for the missing everything and the waiting around for each thing to be passed through all its initial checks and inductions; today was FINALLY the start of my first real experiment!   Using the plates prepped yesterday, this morning was a slightly earlier start in the tissue culture lab to serum starve the cells (which means they have a media containing all the nutrients and antibiotics they need to grow, but without the extra growth factors and signalling molecules (e.g. cytokines) that comes with the BSA, used in the enriched media they have been growing in up to this point). Then we waited most of the day to allow the serum starve to last as long as possible before starting the first timepoint of the experiment - inducing an inflammatory response in the cells with cytokines 24hrs before I assess the response the cells had and find out if they produced the protein I am looking for tomorrow. It's going to be a busy next fe...