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Author Topic: What am I looking at?  (Read 3899 times)

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Offline jfrasca

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What am I looking at?
« on: April 13, 2009, 01:59:08 PM »
Hello All,

   My lab mate just finished some IHC on rat brain tissue. It is perfused and then post fixed with 4% pfa. The sections were cut at 30 microns. In this picture you have TH+ staining and Nissl staining. However, we saw that we have cells with this odd, choppy, nissl staining we have not seen in other tissue. I know nissl staining isnt very specific but could anyone venture a guess to what we are looking at? We will try and H&E later but for the time being we are curious about what type of cell or what type of reaction is occurring in this cell. This system potentially has a lot more mast cells than most rat tissue. The animals were also treated with a neurotoxicant. I am not sure if that really matters since Nissl staining is so non-specific but can any conclusions be made about it?
 
 Thank you,
    Joe

What am I looking at?
« on: April 13, 2009, 01:59:08 PM »

Offline Cardio

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Re: What am I looking at?
« Reply #1 on: April 13, 2009, 03:50:49 PM »
Do you only see it in the treated mice?

Offline jfrasca

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Re: What am I looking at?
« Reply #2 on: April 13, 2009, 04:06:28 PM »
As far as I can tell...Yes.

Offline excalibur

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Re: What am I looking at?
« Reply #3 on: April 15, 2009, 12:41:16 PM »
Without seeing it myself through the scope, it appears to be an artifact on top of your section. Did you filter the cresyl violet before staining?
Paula Keene Pierce, HTL(ASCP)HT
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Offline jfrasca

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Re: What am I looking at?
« Reply #4 on: April 20, 2009, 11:16:30 AM »
Yes, the cresyl was filtered. I believe the consensus is it is an artifact. However, I have not determined if it is a bacteria problem, mounting problem, or any of the other steps it could have been. If I ever figure it out I will post again. Thank you all for your help.

 

Offline Cardio

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Re: What am I looking at?
« Reply #5 on: April 20, 2009, 02:09:46 PM »
What did your H&E reveal? If it is a bacteria problem wouldn't you be able to see it on a Dapi stain? It seems very confusing that you see it only in the treated mice. I would expect a reagent problem or processing issue to show itself in all samples.

Offline jfrasca

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Re: What am I looking at?
« Reply #6 on: May 04, 2009, 03:46:54 PM »
The tissue was from another tech in my lab. I will not know the results of the H&E until he makes it a priority to do it. I suspect that it was a filtration error at some point, but I know the cresyl had been filtered. The PB that was used to mount the tissue, before it was counterstained, could have done it. I have seen that before in unfiltered fluorescent stains. The original tech that showed me that used to call them snosauges (they look like lines of sausages). It was reliably eliminated by filtering all reagents beforehand. I will post again if we find it was something different. Thank you.

Re: What am I looking at?
« Reply #6 on: May 04, 2009, 03:46:54 PM »