[3dem] cryo fib milling of grids for Titan Krios

Gilpin, Christopher J gilpin at purdue.edu
Wed Apr 29 12:12:23 PDT 2015


Hi Everyone,
We are about to embark on some cryo fib milling of cells grown on grids with a view to do cryo tomography. The plan is to use a FEI Quanta 3D FEG fitted with a Gatan ALTO cryo system. I have seen data using this technique but using e.g. a Polara  (Peijun Zhang). Has anyone tried to fabricate holders to take FEI Titan cartridges into a cryo FIB (and out again) and successfully load them into the cassette. We will need to mill the cartridges to allow for a shallow milling angle but that has been shown before by Peijun and colleagues.
I have concerns about the delicate handling of the cartridges to get them in and out of a shuttle system and then into the cassette. Autoloaders are not tolerant of any excess ice.

If anyone has experience or thoughts I would be most grateful to hear from them.

Regards

Chris

Christopher J. Gilpin Ph.D.
Director, Life Science Microscopy Facility
Purdue University
Whistler Hall of Agriculture Research, Room S052
170 S. University St
West Lafayette, IN 47907
765-494-7750
gilpin at purdue.edu<mailto:gilpin at purdue.edu>

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