Chris Tully wrote:
> Anchal,
>
> Regardless of which software you are using, you need some way of
> isolating the ER and Golgi from the rest of the cell.  You may want to
> consider fixing the cells and using a membrane stain for isolate the
> ER and Golgi based on shape, or you will need to express a third FP in
> one of the proteins that populates the ER and Golgi membranes.  With a
> seperate channel it should be relatively easy to derive ROIs from the
> membrane channel for use in measuring the Protein A distribution in
> the main image.  Now, this technique is only partially accurate. 
> Since both the ER and Golgi are relatively small bodies that do not
> fill the entire vertical space of the cell you really need to look at
> this problem in 3D.  You will need a lot of slices because you want to
> get as close to a cubic voxel as possible.
>
> I am sure there are some 3D tools for Image J but I would recommend
> that you also check out commercial software such as 3D Constructor
> (for Image-Pro Plus; http://www.mediacy.com <http://www.mediacy.com>),
> Volocity (http://www.improvision.com/products/volocity/), and Imaris
> (http://www.bitplane.com/).
>
or the newer open source packages, Endrov, OME and Bioimagexd

/Johan

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Johan Henriksson
MSc Engineering
PhD student, Karolinska Institutet
http://mahogny.areta.org http://www.endrov.net