Date sent:              Wed, 02 Aug 2000 11:20:54 -0400
From:                   "Shea Miller" <[log in to unmask]>
To:                     [log in to unmask], [log in to unmask]
Subject:                GFP

The additional fluorescence is almost certainly autofluorescence in
the fungal hyphae.  This is a problem that workers here have had to
deal with.  The obvious test is to look at hyphae of an
untransformed species of the same fungus and see if similar
emmissions are present.

Ian
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> Hello everyone;
>    I have just started looking at a GFP-transformed fungus growing on plant surfaces, using regular fluorescence microscopy.  I can clearly see the fungus using two filter sets: a UV set (ex/em 365/>420) and a blue filter set (ex/em 470/>515).  While the fluorescence in blue light is much
brighter, stuff that is blue/blue-white with UV illumination is green in the blue light.  is the "extra" green in the blue light likely to be where the GFP is not as concentrated, and hence doesn't show up in the UV, or is it more likely to be some other compound that has similar
excitation/emission characteristics?  I have done a fair bit of reading on the subject, but am still feeling a bit vague, and would really appreciate hearing from those of you who have some first hand experience with GFP.
>
> thanks in advance
> shea
>
>
>
> Dr. S.Shea Miller
> Agriculture & AgriFood Canada
> Eastern Cereal and Oilseed Research Centre
> Rm. 2068 Neatby Building
> Central Experimental Farm
> Ottawa, Ontario
> Canada   K1A 0C6
> Phone:  (613) 759-1760
> Fax:  (613) 759-1701
> E-mail:  [log in to unmask]
>
>



Ian Hallett
HortResearch
Mt Albert Research Centre
Private Bag 92 169
Auckland, New Zealand
Fax 64-9-815 4201
Telephone 64-9-815 4200
EMail [log in to unmask]