Acquire seamless images for quantitative TEM
Added by Claudio Schmidli over 8 years ago
Hi,
Do you think it's possible to acquire seamless images over a large areas (e.g. 0.5um x 0.5 um) to do quantitative TEM? Can the alignment of the targets be accurate enough?
I would like to count the amount of protein on a certain area.
Cheers,
Claudio
Replies (4)
RE: Acquire seamless images for quantitative TEM - Added by Anchi Cheng over 8 years ago
Yes with image shift. Is this cryo?
Is 0.5 um the tile size or total ? The reason I ask is that you get 0.4 um size already with 1 angstrom pixel.
RE: Acquire seamless images for quantitative TEM - Added by Claudio Schmidli over 8 years ago
Hi Anchi,
Thanks for the answer. It's for negative stain TEM. Sorry I mean a total size of 0.5 x 0.5 millimeters not microns. The tile size would be about 7x7 microns resulting in ca 5000 images.
So this means with image shift it's accurate enough but I should not use the stage shift?
RE: Acquire seamless images for quantitative TEM - Added by Wim Hagen over 8 years ago
Hi Claudio,
On an FEI TEM, at the magnification you need for a 7 micron tile size, you will only have about 25 micrometer beam-image shift, on some systems just 20, when you have a post-column energy filter maybe even less.
Wim
RE: Acquire seamless images for quantitative TEM - Added by Anchi Cheng over 8 years ago
As Wim suggested, you will need stage shift to get that big an area. Stage shift does come with more error, 1-2 um on a reasonably well maintained one. Therefore you need to add overlap for it.