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Questions from customers

No spots at all

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Hi Mabtech,

 

I received my Human IFN-gamma and TNF-alpha FluoroSpot kit (#FSP-0109-2) on Tuesday and I have just attempted to read the results. 

 

Except there were no spots to read.

 

I think the plate was not activated correctly. I stored the kit box including the plate in the fridge for about 90 minutes before preparing the plate as per kit instructions. The plate conditioned for about 40 minutes before I added my cells and stimuli.

 

Would the 90 minutes in the fridge have prevented the plate from activating correctly? Do you have any other suggestions?

 

I perform about 2 enzymatic ELISpots a week so I am quite comfortable with the concept. However, I have never used pre-coated plates I always activate my plates and store them in the fridge the night before.

 

Kind regards,

J

 

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Hi J,

Sorry to hear about this no spot-issue!

 

The storage of plate in the fridge should not be a problem. To be able to trouble shoot your problem better, would you mind sending over your complete detailed protocol?

 

Just to be on the safe side, I also want to ask if you have used an AID machine for analysis, and in that case if you have switched on the xenon-lamp needed for FluoroSpot (please see attached image)? The external lamp isn’t needed for ELISpot since there is a small LED lamp inside the reader, but for FluoroSpot you need to manually turn on the external lamp. I’m sorry if this is obvious for you, I just had to ask since this solved the issue for another customer just a few months ago.

 

IMG_0387.jpg

 

If you have the lamp on, and you see no spots, what happens if you crank up gain and exposure? You should at least be able to see the contour of the well. If it’s totally black, there might be something wrong with the lamp. When we turn it on, a fan starts and there is bright light leaking out from the box. Do you have the same experience?

 

If all seems good with the machine and the lamp, then as said above, please forward your protocol in detail so that we can have a look at it.

 

Kind regards,
Jens

 

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Thanks for the advice.

 

I did read it on an AID machine but I will go back to it and check the lamp. We did some beta testing on this Mabtech kit late last year.

I used the same profile settings as last time, and we saw spots then. However, I did not turn the lamp on the box and I am completely happy if this is the problem.

 

I have set up another plate and will attempt to read this one and the original on Monday. 

 

Kind regards,

J

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