Equipment

PaCIFiC houses a Leica EM GP2 for vitrifying (plunge-freezing) samples on grids, a Leica STELLARIS 5 cryo-confocal microscope, and a Glow Discharge Cleaning System optimized for cleaning TEM cryo grids, in addition to facilities for biological sample preparation (cell culture and infection with hazard group 2 pathogens).

Leica EM GP2

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The EM GP2 can prepare grids for single particle analysis and vitrifying cells grown on electron microscopy grids for cryo-imaging.

Automatic single- or multi-sided blotting
Environmental control (temperature and humidity)
Viewing system (Leica S9E) for accurate alignment

⬇⬇ Blotting in action ⬇⬇

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STELLARIS 5 Cryo-Confocal

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STELLARIS 5 Cryo is a cryo-confocal microscope system that helps you target your area of interest for cryo-electron tomography (CryoET). The STELLARIS Cryo gives you the precision to target reliably through the LAS X Coral Cryo software that allows for precise positioning of open format coordinate markers on a nanometer scale.

Technical Specifications:

White light laser (485-790 nm) plus solid-state lasers (405 and 448 nm)
50× 0.9 NA cryo CLEM objective (HC PL APO 50x/0.90)
Cryo-CLEM stage with liquid nitrogen reservoir
5 Power HyD detection channels
Tau sense modality
Lightning Confocal Super Resolution modality
Transfer shuttle with cartridges for either bare grids or clipped grids
Enclosed cover box to avoid ice contamination during grid loading/unloading
K5 sCMOS camera for brightfield/reflection/epifluorescence grid mapping
LAS X software (including Coral Cryo)

Glow Discharge Cleaning System

PELCO easiGlow™ is primarily designed for cleaning TEM grids and hydrophilization of carbon support films, which have the tendency to be hydrophobic.
A glow discharge treatment with air will make a carbon film surface negatively charged (hydrophilic) which allows aqueous solutions to spread easily.