Cryo-preparation for TEM, Microtomes

Instruments for sample preparation for cryo-TEM and regular TEM include:

FEI Vitrobot Mk. III

For vitrification of small samples (proteins, protein complexes, liposomes and exosomes) for cryo-TEM imaging.  The sample is applied to an EM grid with a holey carbon support film, excess liquid is blotted off and the grid is plunged in liquid ethane to vitrifie the sample without ice crystal formation. For training and sample preparation services , please contact (979-845-1607).

Leica EM ICE High Pressure Freezer with Light and Electrical Stimulation

Considered the gold standard in sample preparation for TEM. Samples are loaded in cartridges and rapidly frozen at high pressure without ice crystal formation. Samples can be subjected to light or electrical stimulation at defined time before freezing, allowing to capture rapid biological processes, e.g., synaptic vesicle release, or sunscreen degradation upon UV light exposure.  The EM ICE also has cartridges for ells grown on sapphire coverslips, copper tubes for freezing cell suspensions for cryo-sectioning and cryo-TEM tomography (CEMOVIS) and a number of other accessories.

Please contact  (979-845-1607) for further information, training and sample preparation services on this and the AFS2 instruments.    

Leica EM AFS2 with automatic liquid handler FSP

Once the samples have been frozen in the EM ICE, they can be processed (freeze-substituted) for regular TEM by replacing the water by a suitable solvent (acetone) at low temperature without ice recrystallization. Additional protocols include progressive lowering of temperature (PLT).

After freeze substitution, the samples are infiltrated with embedding resin, either epoxy (for structural studies) or acrylic (for immunolabeling). The FSP liquid handler allows automatic solvent and embedding media exchange in protocols that do not use osmium tetroxide.

Leica UC7 ultramicrotome with the FC7 chamber for cryo-ultramicrotomy

For ultra-thin sectioning of resin-embedded samples for TEM. The cryo-chamber allows low temperature ultrathin sectioning of frozen biological samples and sectioning of soft polymers that cannot be properly sectioned at room temperature.

For more information, sectioning services and training, please contact (979-845-5928).

Reichert Ultracut ultramicrotome

For ultra-thin sectioning of resin-embedded samples for TEM.  

Micron Rotary Microtome for sectioning of wax-embedded samples for light microscopy

This microtome is for routine sectioning of biological samples embedded in paraffin wax or Steedman's wax. Sections thickness can be set between 4 and 60 micrometers.  Please contact  (979-845-1607) for further information, training and sample preparation services.

American Optical heavy duty sliding microtome

For sectioning of hard and tough samples for light microscopy. Samples like wood or bone are a often sectioned this way. Please contact  (979-845-1607) for further information, training and sample preparation services.