Freeze-etching

Freeze-etching removes surface ice from a frozen specimen by sublimation under vacuum, often after freeze-fracture microscopy has exposed a new face. Removing that ice can reveal structures initially hidden beneath it. Fracturing opens the specimen mechanically; etching subsequently changes the exposed surface by removing material.^[ref]

A physical preparation holds the specimen cold in a controlled vacuum, permits a chosen amount of ice removal, and then applies the coating appropriate for electron microscopy. The exposed block face can be examined with a scanning electron microscope, or a supported replica can be examined by transmission electron microscopy. Coating and transfer preserve information about a prepared face rather than making the original bulk material transparent.^[ref]

For an own geometrical model, suppose a flat ice surface recedes uniformly at $v=0.5\mathrm{nm}{\mathrm{s}}^{-1}$ while an embedded feature does not sublime. A feature initially $d=10\mathrm{nm}$ below the surface is first exposed atAfter $40\mathrm{s}$ the ice has receded $20\mathrm{nm}$, leaving the feature protruding $10\mathrm{nm}$ in this model. The numbers are stipulated, not a measured etching rate or a recommended preparation setting.

A useful control compares matched specimens after different etching durations while recording specimen temperature, transfer conditions and coating geometry. Progressive exposure would be consistent with ice removal, but a different appearance can also arise from changed preparation or contamination. Condensation deposits material rather than removing it, so contamination must not be mistaken for an etched feature.

The ideal recession calculation assumes a uniform rate and an unchanged embedded feature. Real local composition and heat transfer can violate both assumptions. Interpret image contrast together with etching and coating history rather than assigning every relief feature directly to the initial specimen.