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Electron Microscopy Sample Preparation

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Optimal sample preparation is key to high quality EM data.  The Imaging Facility has a suite of tools to prepare samples for electron microscopy including a high pressure pressure freezer to preserve optimal ultrastructure and a freeze substitution unit for automated sample dehydration.  The Facility has a dedicated workspace for chemical fixation, heavy metal labelling and resin-embedding.  For microtomy there is a semi-automated ultramicrotome used to cut ultra-thin sections and serial ultra-thin sections for array tomography.

 

Our Systems:

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Leica Artos 3D Ultra Microtome

Semi (0.2 - 2 碌m) and ultra-thin (50 - 200 nm) sections of resin embedded material for EM analysis

Trimming of resin samples for FIB-SEM

Array Tomography

Active damping platform with a dynamic range of 0.7Hz to 1kHz, passive beyond 1kHz

Array tomography capabilities including AT4 knife

Diamond trimming and sectioning knives

M80 stereo microscope and colour digital camera

Leica EM ICE High Pressure Freezer

Cryo-fixation and vitrification of specimens up to 200 碌m in depth

Used prior to freeze substitution with Leica EM AFS2

Automated specimen loading

Cryo-fixation in < 1 second

No loss of ions/molecules or lipids

No denaturation of enzymes

No osmotic effects

Preserves antigenicity

Specimen holders for CLEM, tissue biopsies, suspensions and small organisms

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Leica EM AFS2

Automated Freeze Substitution (AFS) of high pressure frozen specimens

Progressive Lowering of Temperature (PLT)

Automated liquid handling with the Leica EM FSP (freeze substitution processor) 

Efficient UV polymerization with built-in LED UV lamp

Minimizes operator contact with toxic material

Reduces risk of specimen loss

AFS sample holders for up to 20 specimens

Compatible with manual embedding in cryo vials

Leica EM CPD300 Critical Point Drier

Critical point drying of specimens for SEM imaging

Fully automated programmable process

Specimen holders for coverslips, small organisms, suspensions and gels

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