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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 Crescent-like development of well
Date
Wed Dec 19 2007 17:11
Author Kim Z
(kzinnack@therionbio.com)
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We are new to this elispot procedure...we have been noticing that we get non-uniform development of our wells, a cresent like appearance of spots that does not happen in every well. We are using a protocol for murine IFN gamma with mouse splenocytes. It almost appears as if we are getting air bubbles in the wells (but upon inspection after reagent/sample addition there are none). All sample/reagent additions and washes have been done with a multi-channel pipettor, however the areas of non-development do not always occur in the same place of the wells within the plate. We have taken the posted advice and dried our wells immediately after development/washing, so it is unclear whether this is the cause. Are the washes too rigorous? Does anybody have any advice for such a problem? We have also performed an ELISPOT kit (IL-2) by R & D systems that recommends incubating the plates/ the bottom molded in aluminum foil ("should minimize the edge effect"), would this help? Would appreciate any advice... Thanks!
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