A method for monitoring the collapse of plastic sections as a function of electron dose.
We present a method for monitoring the collapse of plastic sections when irradiated in the electron microscope. The two surfaces of the section are separately coated with colloidal gold particles. The section is then tilted to an angle of 45 degrees in the microscope and a series of micrographs recorded, corresponding to increasing total electron dose. The collapse of the specimen normal to the plane of the section causes a relative movement in the image of the two sets of particles marking the two surfaces. By measuring the positions of a few gold particles on each side of the section in each exposure of the series, the collapse and also the in-plane shrinkage can be computed. The sections exhibit a rapid initial collapse, followed by a much slower phase of thinning. These effects should be taken into account when producing quantitative three-dimensional maps from tilt series of sectioned material.
Luther
P K
PK
Biophysics Section, Blackett Laboratory, Imperial College, London, UK.
Lawrence
M C
MC
Crowther
R A
RA
D000818:Animals; D004330:Drosophila; D008854:Microscopy, Electron / Q000379:methods*; D009132:Muscles / Q000648:ultrastructure; D010969:Plastics; D014335:Tropomyosin
D010969:Plastics; D014335:Tropomyosin