Hi guys,
I would like to know how to properly use the Pefect Loop in order to pick up ultrathin sections and release them well oriented onto a formvar slot grid.
Thank you very much!!!
Francesco
"... use the Perfect Loop to move single ribbons to a coated slot grid.."
Citation and description of "Picking up sections" from
Anatomical Methods Homepage
Serial Thin Sectioning - The Hartwieg Method
by Hall, Hartwieg and Nguyen
citing:
Polishchuk, R.S., Polishchuk, E.V., Marra, P., Alberti, S., Buccione, R., Luini, A. and Mironov, A.A. 2000. Correlative light-electron microscopy reveals the tubular-saccular ultrastructure of carriers operating between Golgi apparatus and plasma membrane. J. Cell Biol. 148: 45–58. ( http://jcb.rupress.org/content/148/1/45.full )
cf.: EMSTSFIG 3 (within text= small, at bottom of page: big sketch: "Thin section pick up by the Hartwieg method. " found in: http://www.wormatlas.org/EMmethods/STSHartwieg.htm
Best wishes and good luck!
Dear Francesco, you're welcome....and naturally I do hope you're satisfied with the method shown there....from my own experience in "former times" when I needed to position 3-4 serial sections unto formvar-mounted TEM-(classical mesh-)grids I know that these manual movements of the hand will take time to be +/- automated yielding everytime good final results.
Dear Francesco,
In addition to the suggestions from Wolfgang, I've personally had good experiences with the following method (though it requires quite stable hands):
pick up a formvar slot grid (with glow-discharged carbon coat) with N5 (crossover tweezers) and hold it i your left hand (or right hand if you're left handed). pick up the ribbon with the perfect loop.
now tilt the loop towards you while looking through the binoculars of the ultramicrotome
and you should be able to see the ribbon in the perfect loop.
approach with the slot grid to the upper side of the perfect loop (film facing the droplet with the ribbon) orient the slot of the grid parallel to the ribbon and touch the droplet with the grid. (see picture) This should transfer the ribbon to the grid.
best wishes and good luck!
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