Egfp thin layer

[Daniel James White]

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Hi J-Y

Confocal or widefield?

Widefield will see not much difference in image or mean intensity if the layer is in or pit of focus as there is no contrast in xy direction

In confocal you will see a slice that looks like the psf integrated continuously over xy 

Anti gfp antibodies coating the coverslide ( 0.17 exactly mm of course) will work if you can get pure egfp. You could ask the protein purification facility at mpi-cbg. They have lots. 

What's the assay and why? 

You know why I'm interested! 

Dan  

On 21.09.2010, at 07:01, CONFOCALMICROSCOPY automatic digest system <[hidden email]> wrote:

Dear friends and colleagues,

For a microscope calibration assay, I am trying to image a thin layer =20=

of eGFP-like fluorophore.

I aim at imaging a thin layer of fluorophore deposited on a microscopy =20=

slide. I turned to the Murray and colleague solution (1), but they =20
image fluorescein, which spectra are quite different from eGFP.

I have been looking for eGFP preparation commercially available, but =20
unfruitfully so far.

The closest I could find was the BD Living Colors EGFP Calibration =20
Beads, but these beads are 7-10 =B5m in diameter, larger than the depth =20=

of focus of the systems I will use.

Would you know of any solution to get or purchase eGFP on small beads?

Thank you very much
Best regards
jyt




(1) Murray et al. Evaluating performance in three-dimensional =20
fluorescence microscopy. Journal of microscopy (2007) vol. 228 (Pt 3) =20=

pp. 390-405



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Daniel J. White Ph.D.

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