I use a special tool for this, The perfect loop. I do this as follows:

The ultra thin section is picked up with this perfect loop (I have no affiliation to EMS, we just paid for it ;). I fit the handle of the loop under a stereo microscope in a self-made holder. This is in fact a tube holder with many holes. The top of the handle is laid on a polystyrene rod, the one used for cleaning a ultratomy knive. The loop is now horizontally.

The single-slot grid w/ formvar coating is picked up with a tweezers. Bring it to the bottom of the loop. When you come near, the drop of water in the loop will pick up the grid.

Filter paper is used to draw away most of the water. I use the sharp angle of wedges cut from round 90mm black ribbon filter paper. During this procedure the section is kept in the middle of the grid with a hair. I use hair from guinea pig (Cavia porcellus) on top of a bamboo stick. The last bit of water must evaporate.

Check with the microscope if all the water is evaporated and now pick up the grid.

I now other groups use another tool to do this. Now it is another EMS tool: EMS Domino Rack

The formvar coating is put on top of this rack. The ultrathin section is picked up in the single slot grit without formvar, the transferred to the rack. Placed on top of a hole and left to dry. I want to try this method soon.

I do it the same way and it can be frustrating for me as well. I sometimes make a few extra sections, tilt the grid under water and collect the extra sections with the top edge of the grid, so that the rest of the ribbon ends up on the slot. This is good enough for my needs, but if you want a continuous series of sections on several grids without missing a single one, it isn't a good way to do it, I guess.

Other people collect from the top - they press the grid on the surface from above and collect the sections that way. I have no idea how they know they got it right, but they pull it off really well. I can only be amazed, though.

I use a special tool for this, The perfect loop. I do this as follows:

The ultra thin section is picked up with this perfect loop (I have no affiliation to EMS, we just paid for it ;). I fit the handle of the loop under a stereo microscope in a self-made holder. This is in fact a tube holder with many holes. The top of the handle is laid on a polystyrene rod, the one used for cleaning a ultratomy knive. The loop is now horizontally.

The single-slot grid w/ formvar coating is picked up with a tweezers. Bring it to the bottom of the loop. When you come near, the drop of water in the loop will pick up the grid.

Filter paper is used to draw away most of the water. I use the sharp angle of wedges cut from round 90mm black ribbon filter paper. During this procedure the section is kept in the middle of the grid with a hair. I use hair from guinea pig (Cavia porcellus) on top of a bamboo stick. The last bit of water must evaporate.

Check with the microscope if all the water is evaporated and now pick up the grid.

I now other groups use another tool to do this. Now it is another EMS tool: EMS Domino Rack

The formvar coating is put on top of this rack. The ultrathin section is picked up in the single slot grit without formvar, the transferred to the rack. Placed on top of a hole and left to dry. I want to try this method soon.

There really is only one way and that is to hold your grid in fine forceps and gently enter into the water the sections are floating in .  Drop underneath the sections and slowly raise up under the sections with your  grid,  Blot of any water and let the grid dry. It dries typically very quickly.  The alternative is to approach from above the sections and touch  the grid to the sections.  If you add a little extra water the sections will be easier to touch.  Either way works well with training, but no coffee on that day!  Once dry, stain with heavy metals (wash well between metals).  Make sure your grids have been cleaned well before use.

Hi Ian,

I use self-closing tweezers, hold the slot grid submersed so that only a tiny bit of the slot is visible outside of the water. I then guide the ribbon towards that water edge and also use a hair tool to manoeuver the ribbon in the water (the quality of the hair tool is quite important, if it gets sticky the sections will stick and you will easily loose precious sections. Keep it clean and try to use elastic, thin hair, e.g. thin eye lash - I use the coating fur of a German shepherd ).

Once the first section of the ribbon makes contact with the water edge it will stick to it and I simply slowly lift the grid out of the water and the ribbon will nicely flatten in the slot. It all depends on how the ribbon is oriented while it makes contact with the water edge. Your problem might be that you do not properly align the ribbon with the slot before lifting the grid. Try to practice the attachment of ribbon to water edge as this is the crucial step to get the ribbon nicely placed inside the grid. For me it works quite well. After I lift the grids out of the water I leave them to dry while still in the self-closing tweezers to prevent ripping the film (I use 1% pioloform in chloroform which is quite forgiving in terms of handling). I do the same after the washed following the heavy metal contrasting.

I would not recommend pressing onto the section with the slot grid from above as you have no control over how the sections will be placed inside the grid and the risk for folds in the sections is increased dramatically.

Hope that helps, the best tip is simply to practice, practice, practice ;-)

Best wishes and good luck!

Christian

Check out the method that is just about foolproof, though laborious at:

W.H.Fahrenbach (1984) Continuous serial sectioning for electron microscopy.

 J. Electr. Microsc. Technique  1, 387-398

Thank you all for taking the time to answer my question. I'm sorry to have taken so long in getting back to you all but I have been vigorously refining my technique on the ultramicrotome since then. I am still using my traditional method of submerging my coated, slot-grid and guiding with my hair tools I made from the coating fur of beagles.

Since this time I have become much better at getting the sections on and flattened to the PEI coating. I have noticed that cleaning my hair tools helps quite a bit so thank you to Christian. 

I made my own versions of the perfect loop tool using wire and pippette tips as my handle as well. I find this great for mounting thick sections to glass slides for Toluidine Blue staining but have not had great success with it in terms of mounting thin sections.

I have read some about this domino technique but it still seems a bit odd to me.

Wolf, thanks for the reading suggestion. I have read this paper before but I may need t have another look.

Again, thank you all for your help. I really appreciate it.

Ian 

Hi all, 

I thought I would update on this process given the interest this thread has received. I had always used the technique I spoke of previously on this thread (submerge and coax method) but had run into problems and was looking for a solution. 

During this time I experimented with the "perfect loop tool", a variety of grid types and "plucking" sections from above.

Since, ordering new copper-beryllium slot grids and not recycling/washing old grids, my difficulty lessened and then finally, my support film solvent, dichloro-ethane had gathered too much moisture and left me with inadequate/dirty grids. Once I ordered new solutions and prepared new coated-slot grids most of my trouble went away when using the submerge method.

Along the way, I worked with varying mesh-grids during the pilot part of my imaging study and found that using the perfect-loop tool was fantastic when using the mesh grids without support film. However, when I tried using the perfect loop for any of my coated grids it often resulted in a bursting of the support film or improper placement of the sections on the grids.

I also found that the "plucking" method used to dab the surface of the water bath where sections are worked pretty well for most grid varieties coated and non-coated alike. The issue with this technique was that it lacked consistency when picking up one ribbon of sections to the next in terms of their location on the grid and the degree of wrinkling observed on the sections.

I hope this helps. Cheers,

Ian

I also use a singly, eyelash taped to a small bamboo stick. Holding the grip in one hand and carefully guiding the ribbon to the grid by use of the eyelash. I hope this helps!

Best