Figure 1 | Scientific Reports

Figure 1

From: Cryopreservation of infectious Cryptosporidium parvum oocysts achieved through vitrification using high aspect ratio specimen containers

Figure 1

Design of high aspect ratio vitrification cassette. (a) The vitrification cassette consists of two sheets of polycarbonate laminates (thickness 178 µm) bonded by a sheet of pressure sensitive adhesive (thickness 120 µm) which is cut out to form the chamber. Outer dimensions are 52 mm by 40 mm, with internal volume of ~ 200 µL. (b) Solutions are loaded into the cassette via pipette tip affixed to a PDMS punch (4 mm outer and 1 mm inner dimeter), which seals the cassette port and creates a vacuum. (c) Formation of vitreous or crystallized state is determined by visual inspection. Images reflect contents of cassettes at × 20 magnification immediately upon removal from liquid nitrogen. Pure water appears opaque indicating ice formation. A solution of 25% DMSO appears to vitrify, though ice formation occurs during transfer from liquid nitrogen to the microscope stage appearing as opaque blotches in the image. 50% DMSO appears to vitrify, though cracks form during the transfer from liquid nitrogen to the microscope stage. Supplementary Video S1 shows time-lapse of melting.

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