New controlled environment vitrification system for preparing wet samples for cryo-SEM

A new controlled environment vitrification system (CEVS) has been designed and constructed to facilitate examination by cryogenic scanning electron microscopy (Cryo-SEM) of initial suspension state and of microstructure development in latex, latex-composite and other coatings while they still contain solvent. The new system has a main chamber with provisions for coating as well as drying, and for well-controlled plunging into cryogen. An added subsidiary chamber holds samples for drying or annealing over minutes to days before they are returned to the main chamber and plunged from it. In the main chamber, samples are blade-coated on 5 × 7 mm pieces of silicon wafer and held at selected temperature and humidity for successively longer times, either there or after transfer along a rail into the subsidiary chamber. They are then placed in the sample holder mounted on the plunge rod, so as to permit adjustment of the sample's attitude when it plunges, at controlled speed, into liquid ethane at its freezing point, to a chosen depth, in order to solidify the sample without significant shear or freezing artifacts. The entries of plunging samples and related sample holders into liquid ethane were recorded with a high-speed, high-resolution Photron digital camera. The data were interpreted with a new hypothesis about the width of the band of extremely rapid cooling by deeply subcooled nucleate boiling below the line of entry. Complementary cryo-SEM images revealed that the freezing rate and surface shearing of a sample need to be balanced by adjusting the plunging attitude.