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Hi Monuique,
First you want to get rid of the air that is in the intercellular spaces in the 
leaf. As George Littlejohn mentioned, perfluorodecalin is one option. One small 
downside in my hands is that there are always some small air pockets left 
every now and then (usually in the most interesting area of the leaf).  
Often, I just vacuum infiltrate a leaf segment with water, using a syringe 
(works much better than a a vacuum dessicator: put the leaf in a 10 ml plastic 
syringe, add about 1 to 1.5 ml water, insert the plunger and push all the air 
out of the syringe. Then put your finger on the syringe outlet and pull the 
plunger to creat vacuum. Shake the syringe vigorously to disodge air bubbles 
sticking to the leaf. When the leaf is under water, release the plunger. The 
water gets drawn into the leaf, the leaf should become translucent. If needed, 
repeat few more times.    
Once infiltrated, I mount the leaf in water in a coverglass-bottom chamber (we 
have an inverted microscope) - either a standard coverglass-bottom Petri 
dish, or my home-made plexiglass chamber - 
http://microscopy.tamu.edu/lab-protocols/light-microscopy-protocols.html

I then use a glass "brick" to keep the leaf flat - a piece of glass for making 
glass knives for resin sectioning. It is a good idea to dull the edges of the 
glass block with sandpaper, otherwise you will be cutting your fingers.
     
If you have the leaf infiltrated, the production of gas during illumination is not 
noticeable (I do not see any new air bubbles in the infiltrated areas) 

Sometimes, when in a hurry, I do not bother with vacuumm infiltration of 
Arabidopsis leaves. i just put the leaf in the coerglass-bottom chamber in a 
drop of water, put the glass brick on top and tap with the back of my 
tweezers to knock out the air. Not perfect, but you can always find an area 
without too many air bubbles. The downside is that it is fairly easy to break 
the coverglass bottom by tapping. 

Good luck!

Stan Vitha
Microscopy and Imaging Center
Texas A&M University

On Tue, 24 Jan 2012 15:31:42 -0600, Vasseur Monique 
<[log in to unmask]> wrote:

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>
>Dear all,
>
>Do some of you could suggest how to prepare live sample of plant leaf for 
>confocal microscopy.  We don't have much idea how to "sandwich" a leaf 
>between slide and coverslip so that it is flat et not too thick for microscopy 
>(and without destroying it)? What should we use as mounting media? What 
>should we be aware of?  Our plant will have YFP in chloroplastides.
>
>Thanks a lot in advance to all of you.
>
>Monique