10 March, 2022
In January 2020, I started my first postdoctoral role within the Sharpe lab at the º£½ÇÉçÇøÂÛ̳. I was asked to write a blog post about a typical day in the life of a post-doctoral researcher. Although this sounds simple, as soon as I put pen to paper I quickly came to realise a typical day does not exist. Despite planning, each day undoubtedly brings up new challenges but also new opportunities. Hopefully, I can give you some insight into what I aim to do in the lab each day and the bigger picture for what we are all working towards as a group.
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Together in the Sharpe lab, we aim to understand how a family of proteins, tyrosine phosphatases, mediate the cell’s behaviour in response to local environmental changes such as exposure to growth factors and oxidants. Tyrosine phosphatases catalyse the removal of phosphate groups from tyrosine residues of substrate proteins. The removal of phosphate groups by phosphatases and addition by kinases, mediates reversible conformational changes in the substrate protein’s structure, modifying its function. Functional modifications of proteins can be part of normal, healthy signalling pathways within cells, however they can also be distorted in diseased states. It is therefore important for us to gain fundamental knowledge of the roles that tyrosine phosphates play across cellular signalling pathways in healthy cells, to further improve opportunities for therapeutic intervention when diseased states arise.
Each of our projects are focused around a different phosphatase and our goals are to identify its interactors, involvement in signalling pathways and how external influences can affect normal signalling. I predominantly use cell-based and in vitro assays to answer these questions, focused around my phosphatase of interest, PTPRK. Phosphatases are relatively understudied compared to their counterpart kinases but are shown to have key functions across a range of diseases, therefore being crucially important and interesting proteins to study. This is particularly why I enjoy my work – we really don’t know what we are going to find!
A typical day in the lab
A typical day starts around 7:00 after snoozing multiple alarms and hearing the crash of the dog as she’s been released upstairs for my final wake-up call. After a quick cuppa, slice of toast and dropping the boyfriend off at the office (the shed), I drive into work. At around 9:00 with another tea in hand, I start on the emails and checking my lab book for the days plan. My work in the lab encompasses the design, execution and analysis of experiments. On the back of experimental work, I also contribute towards the wider picture; sharing, conversing and collaborating with both internal and external groups and facilities. Despite planning and preparation, most days don’t go entirely to plan and every day is inevitably going to be completely different. As a scientist, this is part and parcel of the job and enables us to become excellent problem solvers. The challenge of constantly rethinking experiments and trying out new hypotheses is what really excites and motivates me in my job.
After replying to any urgent emails, my first lab job usually involves my cell-based work or what we call tissue culture. Cells are the work-horses of the majority of my experiments so I would usually either be maintaining them by refreshing their culture media or expanding them for experiments. If, on a typical day, my cells were at the stage of being ready for an experiment, I would instead be treating them. This can include treating with different inhibitors, oxidants or growth factors at range of concentrations and time points depending on the designed experiment. After treating for the desired amount of time, I would then finish the experiment by lysing them which is essentially breaking open the cells.
After tissue culture, a big part of my day then involves carrying out techniques to analyse the results of my experiments. My bread and butter technique for analysis is Western blotting and often features in my typical day. Western blotting is a method which allows us to visualise the levels of specific proteins among a mixture of thousands from our cell-based experiments. By looking at changes in protein levels, this enables us to understand how our cell treatments can alter signalling events in relation to our particular proteins of interest, helping us to draw our conclusions.
Western blotting is just one technique that we use in the lab to analyse our experiments. We also perform activity assays, pull-down experiments and make use of our on-site facilities and expertise in using flow cytometry, mass spectrometry and microscopy to name a few. Having the opportunity to use a range of techniques to analyse our experiments is incredibly useful.
By around 12:30, its lunchtime. We try as a lab to have lunch together when we can. In the warmer weather that means sunning it up on the benches outside the building (bring on the summer) but if not then it’s to the canteen instead. I really enjoy taking this time to chat to colleges as it breaks up the day and helps to reenergise for the afternoon. If it’s a really busy day though, it’s usually lunch at the desk instead.
From 13:00 onwards, I usually consolidate everything from the morning’s activities. This involves completing experiments and Western blots, processing samples for the following day’s analysis and also collecting results. I try to spend the last hour or so at my desk to record the day’s progress in my lab book and try to plan for the following day. Another really important aspect of my work is to keep up to date with current literature around my project. I usually get email notifications to update me with new releases but amongst our lab, we send each other papers that might be of interest to the group too.
Although everything I have spoken about so far in a typical day is mainly performed individually, my day also involves a lot of collaboration and communication too. Within our lab, we are constantly receiving and passing on tips for particular techniques, getting to know each other’s projects whilst discussing results and conclusions. We have regular lab meetings that help us to resolve experimental technical issues but also enable us to think about and discuss the bigger picture of our projects too.
Using the expertise of our Babraham facilities staff for experimental advice and also communication with external collaborators for particular projects is another big part to my day. This ultimately enables my work to progress as quickly and efficiently as possible. With an increased capacity and acceptance for using online platforms for communication, this has made it incredibly easy to keep up to date with projects that span across different groups, by enabling us to continue with Departmental Signalling meetings throughout the COVID pandemic. This opportunity has been invaluable for advice on our own projects in addition to increasing our knowledge and insight into what other groups are working on. Although we talk about our projects as personal things, there is a whole team of people behind each one and its really important for me to dedicate some of my day towards this.
Keeping a balance
Once all my lab work is completed and my plans for the following day are set, I head home for the evening at around 17:30. For me, it’s incredibly important to have a really good work-life balance. If I don’t give myself the time to relax, lab work can become a little overwhelming so I actively plan to get involved with things outside of work to help me to switch off. Once I get home, we usually take the dog out for a walk or run around the village. Its great to actually be forced to go outside no matter what the weather after usually being at the bench or desk all day. As well as running, I also play hockey with a fantastic local club. I think this really helps to unwind and get ready for the following day in the lab.
10 March 2022
By Katie Mulholland