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Elispot Discussion
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Please, feel free to start discussions about the Elispot technique, methods, ideas, questions, experience etc. With your input, we would like to make this website to a special interactive place for everyone who is interested in the Elispot assay.
We apologize for not having this discussion site online for a while. Our web site was targeted by someone who placed a bad activeX link in the discussion. We are in the process of recovering as much as possible. The shown placement dates do not reflect the actual message posting. The first discussion topic was placed in 1999.
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Subject spot distribution
Date
Wed Dec 19 2007 17:12
Author Sylvia
(sylvia@zellnet.com)
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Hi Kim;
I believe that the crescent-like development of the wells can be avoided
by doing the following:
1) If your whole well is developed (stained), but all of your spots are
in one area, the cells are added to vigorously to the plate. Please,
be very gentle when adding the cells. Otherwise, they roll towards the periphery in the direction
one is holding the pipette tip with the center of the pressure forming
the crescent distribution. Let the cells settle for 1-2 hours, before you
are adding the antigen-presenters.
2) If a part of your well is not developed at all, it might be due to
incomplete coating. This can happen in PVDF plates because of their hydrophobicity. Best way to
avoid this is to prewet the plates. You find a protocol for prewetting under our
protocol section.
The drying of wells after spot development will not cause this
spot distribution effect.
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