When developing my 4 colour fluorospot plate, i noticed that there was patchy ring of red on the edges of my well. My analytes were IFN-Y, IL-2, IL-17A and TNF-A. I was wondering what this could be?
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Patchy red ring on the edge of wells on fluorospot plate
- GGGuest Guest2024-07-16T15:28:34Z
- aadmin2024-07-17T07:56:39ZAdmin
Hi,
When analyzing cell samples that contain a lot of apopototic cells you can sometimes end up with artefacts in the outer edge of the well. The reason why is elusive, but I believe it is caused by a natural tendency of debris to get pushed out to the edge during the natural course of the experiment and detection phases. Some antibody detection systems are more prone to getting stuck in the debris and you therefore observe the phenomenon more in some cytokines than others.
There are ways to handle this: avoid low cell viability, wash alot during detection and reduce the size of the AOI (area of interest) within the analysis software of IRIS.
Another source of problem could be issues with leakage. It is a bit hard to say from just the single picture you posted. Please comeback with larger image, brightened showing only the affected filter. I can tell you more after that.
