I got the splenocytes from mice spleens and I did ELISpot for the first time. Unfortunately, something had gone wrong and I didn't get the spots. I stored the remaining splenocytes at 4C. Can I use these stored splenocytes to redo the ELISpot?
No, that's too long. I'm afraid that you'll have to do the whole experiment again. At best you can keep the splenocytes overnight at 4oC (that's OK for FACS, probably not so good for ELISpot).
I assume your splenocytes were stored in a tube rather than in a plate. Was it in culture media? If so, some might still be alive (check under a microscope), but they won't produce very nice spots or give you correct proportions of secreting cells as many of the cells will have died. If you're just practising ELISpots, see if you can get hold of some hybridomas to play with.
You should practice with your positive control .I have done lot of Elispots and always do control wells. You can use ConA for mouse splenocytes as positive control.
Not a good idea. Most of the cells will be dead or in very bad shape.
I agree with previous author and need to verificate dead or live cells
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