What optimization steps did you conduct when starting your automated processing?
This question is part of the following Ask The Expert session:
How to get the most out of automating your ELISA-like assays, dos and don’ts
Job Title: Senior Researcher for the High Throughput Antibody Discovery team
When optimizing an automated method there are 3 questions you should be asking, in my opinion:
- Are all of the components functioning properly? – This is usually reserved for the liquid handling part of a system. Accurate QC and preventative maintenance are key. You should run liquid handling QC on your instrument at least twice a year. This can be something as simple as dispensing a fluorophore from a reservoir to an assay and reading it to ensure equal signal across the whole plate. Additional steps can be taken to assess relative inaccuracy, such as, if I program a 5 µL dispense, am I getting 5 µL? Any preventative maintenance or calibration that can be done on components should be done and done regularly.
- Are the steps being performed correctly, as programmed? – Check and double check your method steps. Ensure that the right samples are going into the right wells; the right source is going to the right destination; the right plates are being used at the right time. I usually accomplish this by setting up several ‘wet runs’ with several different colored water batches. I will run a single plate to ensure that the steps are being done correctly and in the expected order. I’ll then increase the number of plates being run to see the effect on the overall time-to-completion.
- Is this the most efficient resource allocation? – Resource allocation is the overall driver of method duration in an automated environment. If you write your method in such a way as to have one piece of equipment doing the majority of the tasks, then your method is going to take a long time. Be open to changing the order of steps. A simple change like this may greatly affect your allocation time of a particular resource, and by extension, the overall length of you method run. Also, be open to bracketing time points. If you define a 1 hour incubation +/- 5 minutes and end up with an unacceptable protocol length, try varying the +/- time value. If you set limits in a method step, the software will obey those limits. If it can’t fit a step into that limit it will wait until it can, extending the overall duration of your protocol. It may happen that increasing that value to +/- 7 minutes may be all the adjustment needed.