However, untreated protein crystals often crack if flash-frozen; therefore, they are generally pre-soaked in a cryoprotectant solution before freezing.

In addition to overcoming the inherent fragility of protein crystals, a number of environmental factors must also be overcome.

Nevertheless, in some cases, protein crystals were reported to be perfect enough to achieve images of single dislocations.

Recovery of the protein crystals requires imaging which can be done by the intrinsic fluorescence of the protein or by using transmission microscopy.

Protein crystals are scooped up by a loop, then flash-frozen with liquid nitrogen.