Re: Mowiol and 3D imaging

Posted by Patrick Van Oostveldt on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Mowiol-and-3D-imaging-tp5845120p5848851.html

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Dear,

You also have to take into account that the refractive index of the  
mounting medium can be changed due to solidification.
An increase in RI will induce measuring errors cfr: Visser and  
Braekenhoff some years a go in J.Microscopy and the measured thickness  
will be reduced.



Greetings

Patrick Van Oostveldt

Quoting "Mario Faretta" <[hidden email]>:

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> Dear Christian,
> the main problem with Mowiol and in general with all the mounting media that
> become solid over time, is that there is a compression flattening cells. This
> effect tend to be obviously worst as time goes on and it is particularly
> dramatic in cell monolayers (I tried just for fun to measure nucleus  
> thickness
> after 1 week and more than two weeks: staining was perfectly maintained but
> cell thickness reduced of more than 50% in 1 week and goes down up to few
> microns at the end (at least at the concentration we use)). The point is that
> if you need to use confocal generally you do it for a 3D analysis but if you
> prepare the sample this way you make it 2D. Depending on the goal of the
> experiment Mowiol can be deleterious almost immediately: I remember an old
> pubblication in Cytometry from Tom Jovin (hope I am correct) showing apical
> and basal membrane fusion at the cell periphery due to mounting media induced
> compression...
> hope it helps
> Mario
>
>  Christian Wilms ([hidden email]) wrote:
>  >
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>  > To join, leave or search the confocal microscopy listserv, go to:
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>  >
>  > Dear List,
>  >
>  > I've read that Mowiol is not considered well suited for 3D imaging:
>  >
>  >
>  > Unfortunately, the authors give no specifics on why that should be the
>  > case and I also can't find anything on the web. Does anyone have
>  > details on this?
>  >
>  > Thanks,
>  >
>  > Chris
>  >
>



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