Stereoscopy by tilted illumination in transmission electron microscopy.
We describe a new technique for stereoscopic observation in transmission electron microscopy, employing tilted illumination. A triple-hole objective aperture is used so that the bright beam can pass through with or without tilt. Stereo views can be acquired by tilting the illumination such that the bright beam passes through a pair of symmetrically arranged apertures alternately. The advantages of this technique as compared to the commonly used method of single-axis tilt are: (i) greater speed, potentially at or close to video rate so that live 3D observation is possible; (ii) elimination of specimen movement associated with stage tilt; and (iii) perspective views corresponding to multiple tilting axes which can be realized by installing more aperture-holes and choosing their positions properly. The main limitation is that the angle of tilt is limited by the extent to which the astigmatism, introduced by beam tilt, can be compensated by the objective lens stigmators.
Fan
G Y
GY
Department of Neurosciences, University of California at San Diego, La Jolla 92093-0608.
Ellisman
M H
MH
D000818:Animals; D001001:Anura; D005727:Ganglia, Spinal / Q000648:ultrastructure; D008029:Lighting / Q000379:methods*; D008854:Microscopy, Electron / Q000379:methods*