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Leica EM HPM100, Leica EM VCT100
High Pressure Freezing, Cryo-SEM
Application Note
Superior Cryopreparation of Biological and Industrial
Samples
Commercially available hand creams
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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carrier, withdrawn into an EM VCT100 shuttle then transferred into a Leica EM BAF060.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withdrawn into the EM VCT100 shuttle and then moved to the cryo-SEM.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
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Hand cream with high water content
shows mostly large emulsified pockets of aqueous
material interspersed between few lipid sheets.
Hand cream with low water content shows mostly lipid shee
ts with very small pockets of
emulsified aqueous content.
Samples prepared August 28, 2009 by Kim Rensing Ph.D. at the University of British Columbia,
BioImaging Facility, Vancouver, Canada
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