micrographs - multiframe

MRC

1

(1562, 1670)

SIGNED BYTE

(100.0, 100.0)

Image acquisition;
5 nm voxels: xy pixel set to 5 nm, FIB mill every 5 nm.
SEM: 1.5 kV with 1 nA current, 10 μs dwell time.
EsB detector grid voltage: 1200 V.
Ion beam milling: 30 kV, 700 pA.

Image processing;
Zeiss Atlas 5.2.1.25;
Gradient align
Export of cropped region of interest exported as 8 bit TIF files

Fiji;
Invert
Gaussian blur: Sigma 0.75
Unsharp mask: Radius (Sigma) 1 pixel, Mask weight 0.6
Rotate to straighten XY (D19: no rotation)
Reslice: YZ assigned to XY plane
Scale to 10 nm isotropic voxels

IMOD;
tif2mrc

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