Re: green fluorescent protein

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From:Jamie Erickson <JErickson@genetics.com>
To:histonet@pathology.swmed.edu, Cathie.Crukley@sjhc.london.on.ca
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Cathie,
            I've done a lot of GFP evaluation and the investigator should be able to cryopreserved these samples and then section. Most people have had no luck in processing samples for GFP evaluation, but it can be done. I've formalin fixed samples then cryopreserved, sectioned them and used vecta shield w/ DAPI (Vector Labs)  to evaluate them if the transfection/infection of viral GFP was high enough we saw lots of GFP. Different tissues will be transfected/infected with the GFP-viral construct at different amounts, I think depending on the level of permeability and proliferation.  Liver injected IV lots of GFP muscle injected IM little GFP expression. Go with the cryo it should be a sure thing, also different camera/scopes may or may not see the GFP to the same amount we use a GFP filter set.  Hope it helps


Jamie Erickson
Associate Scientist
Genetics Institute
1 Burtt Rd.
Andover, MA   01810
work : (978) 247-1348
FAX  : (978) 247-1333

>>> Cathie Crukley <Cathie.Crukley@sjhc.london.on.ca> 07/18 11:18 AM >>>

Hi,

I know nothing about green fluorescent protein, but one of our researchers has some retina in formalin that has been labelled with a green fluorescent protein tagged antibody and wants to know what to do with it next.  His intention is to paraffin process and section it and view it under the fluorescent scope.  Or will he have to stain it otherwise?

Thanks,
Cathie

SJHC , London, Ontario






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