Oscar Swindley is a final year PhD student at the Department of Chemical and Biological Engineering at the University of Sheffield. He works as part of the synthetic biology platform development for biopharmaceutical manufacturing team. The lab purchased its first Opentrons OT-2 in August 2019 to automate titre assays for cell culture development—and Oscar runs it.
Opentrons: What kind of research do you do?
Oscar Swindley: I work on genetic and chemical manipulation of monoclonal antibody folding and assembly processes, alongside optimisation and integration of automation procedures.
We have an OT-2 performing a large number of our cell culture operations in our lab. I have just finished our first large screening project where all the cell culture, transfection, sample collection, and titre assays have all been performed on the OT-2.
Opentrons: What’s a typical day in the lab for you?
OS: Currently I am processing the results of our first large scale cell culture screen completed on the OT-2. I have been using the OT-2 to perform a titre assay. The robot supports and streamlines the process. Outside of that, a typical day includes general cell culture, both in shake flasks and plates and other pieces of experiments in the molecular biology lab.
Opentrons: Why did you choose the OT-2?
OS: The purpose of getting a robot in our lab was to improve the reproducibility of our platform. We do high-throughput screening in our lab using 24- and 96-well plates. Our main difficulty was with transfections as when we resuspend the cells at very high density the solution becomes viscous and cells quickly settle. We need to accurately dispense samples across the plate to seed the same number of cells in each well. It’s a really repetitive process with incredibly small volumes of liquids. Our biggest issue is being able to consistently mix and seed the cells as they settle over time.
Opentrons: What specifically do you use the OT-2 to do, and how does it fit into your workflows?
OS: We use the OT-2 to set up and run our 96-well electroporation platform, sample from multiple culture plate formats, and perform plate-based assays. For our electroporation platform specifically, the OT-2 combines the DNA, buffers and cells, before distributing the mixes into the electroporation plate. After transfection the OT-2 dilutes and mixes and seeds the cells before into 24-well culture plates. Additionally, 3 days later we assay the cultures with the robot. This includes mixing and sampling from the culture plates for a variety of downstream assays. Lastly, the OT-2 is able to load samples onto glass slides for our high-throughput cell counter and harvest supernatants for titre analysis.
During a recent set of screening experiments, the OT-2 helped with 22 electroporation plates and associated sampling. It’s great having the robot take only 20 minutes to run a titre assay. I can have a chain going all day.
Opentrons: What was it like to get your Opentrons robot up and running?
OS: It took me about 2 months to write my first protocol script. But once you’ve done one, the information is all there and it’s quite quick to continue to more complex experiments.
I uploaded all my scripts onto my GitHub page, if anyone’s interested.
Opentrons: Had you used any lab automation before your OT-2?
OS: No. I had used a BioMek before with an SOP setup. I learned Python for the first time between August and November to program the OT-2.
Opentrons: Is there anything else you’d like to say about using the OT-2?
OS: It is a good robot, however it doesn’t have any failsafes—i.e., won’t recognise an obstruction or realise if it drops a tip. The problem we’re having now is tracing mistakes when they occur.
Another difficulty was compatibility with 24 well plates, so I had to trick it by taking out every other row of tips. But if you know how to program the robot, you can always find a solution.
Overall, I think it’s really good for research because you can change your protocol every single time.