Leica EM VCT 100, EM HPM100 User Manual

Leica EM HPM100, Leica EM VCT100

High Pressure Freezing, Cryo-SEM

Application Note

Superior Cryopreparation of Biological and Industrial
Samples
Commercially available hand creams

Leica EM HPM100, Leica EM VCT100

Commercially available hand creams

Protocol:

Hand creams having different water contents were applied into the 100 µm cavities of two 3
mm type A sample carriers which were then closed cream sides inwards. The sample
assembly was high pressure frozen with a Leica EM HPM100 and moved to a cooled Leica
EM VCT100 loading station. The closed carriers were mounted on a universal sample
carrier, withdrawn into an EM VCT100 shuttle then transferred into a Leica EM BAF060.
Samples were fractured by pushing the top carrier off with the fracturing knife at -140°C, then
they were high angle rotary shadowed with platinum/carbon. The samples were again
withdrawn into the EM VCT100 shuttle and then moved to the cryo-SEM. They were imaged
with a Hitachi S4700 FE-SEM with the EM VCT100 CryoStage at –125°C.

Summary:
Procedure Reagents Parameters

High pressure freezing 3 mm diameter sample carriers
Freeze fracture -140°C
Coating 5 nm Platinum-Carbon 70° angle to the sample surface

with 90 RPM rotation
Transfer Cooled EM VCT100 Shuttle
Imaging Cryo-SEM at –125°C

Results:

High pressure freezing followed by freeze fracture and cryo-SEM are well-suited for
determining the structure of water-containing industrial materials. Multiple samples can be
processed and examined within a short time with an SEM equipped with a Leica EM VCT100
cryo-stage.

www.leica-microsystems.com