PaCIFiC: Your Gateway to Cryo-Imaging
PaCIFiC is a cutting-edge containment level 2 (CL2) facility, equipped to prepare and analyze both non-hazardous and infectious samples (hazard group 2 pathogens). Our facility offers a comprehensive suite of services, including:
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Vitrification: We can expertly vitrify your samples, a crucial step in the cryo-imaging process that preserves their structure for optimal imaging.
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Cryo-Confocal Microscopy: We utilize advanced confocal cryo-microscopy to generate high-resolution images of your vitrified samples. This vital step allows us to assess sample quality and identify specific regions or structures of interest for further analysis.
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3D Subcellular Targeting: We can generate 3D models of your samples, pinpointing specific regions or proteins for downstream high-resolution imaging techniques like electron cryo-tomography (cryoET) or soft X-ray cryo-tomography (cryoSXT).
- Cell Culture Support: We provide access to dedicated cell culture facilities, including incubators and all the necessary equipment for cell growth and maintenance. This ensures optimal in-house sample preparation for downstream vitrification and cryo-imaging.
What is cryo-imaging?
Cryo-imaging is a groundbreaking technique that rapidly freezes biological samples, essentially stopping them in their tracks, designed to revolutionize our understanding of cells and pathogens. Through the power of cryo-imaging, PaCIFiC allows you to peer into the frozen world of biological samples, preserving them in a near-life state for unparalleled detail.
PaCIFiC is Here to Help!
PaCIFiC is currently under commissioning and will soon be available to researchers. If you are interested in leveraging the power of cryo-imaging for your research, please get in touch with us today. We are here to help you.
Biological Safety
For samples requiring biological containment (up to CL2), a comprehensive risk assessment is mandatory. This assessment will be reviewed and approved by the Department of Pathology's biological safety committee in collaboration with the PaCIFiC facility manager. We will provide guidance and support throughout this process. Please contact us for further information.
Training & Booking
Training is available for all interested users.
Once trained, Members of the School of Biological Sciences at the University of Cambridge (i.e., internal users) can book access via our Booking System. External academic/industrial users can book access by contacting us here to discuss their needs and arrange for necessary training.
Access Charges
Please note, that there are different charging rates for internal vs external users.
- Internal Users: Members of the School of Biological Sciences at the University of Cambridge will be charged at the internal rate.
- External Users: Different charging rates apply to external academic and industrial users.
*The Leica EM GP2 and STELLARIS 5 cryo-confocal access charges include gas usage (ethane and liquid nitrogen). A range of other consumable items are available for purchase separately, including a variety of electron microscopy grids, C-clips and rings, and grid storage units. Please contact us to discuss your specific consumable needs.
*The class 2 microbiological safety cabinet and CO₂ incubator include reasonable use of essential consumables (gloves, filter tips, plastic serological pipettes) but exclude tissue culture dishes/flasks or mediums. Users are required to provide their own or contact us for details on sourcing options.
Contact details
PaCIFiC is currently under commissioning. If you are interested in using PaCIFiC please get in touch with our facility manager Dr. Luay Joudeh lj360@cam.ac.uk and/or with our directors, Professor Colin Crump cmc56@cam.ac.uk and Professor Stephen Graham scg34@cam.ac.uk.
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
⬇⬇ Blotting in action ⬇⬇
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
STELLARIS 5 Cryo-Confocal
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.
Cell Culture & HG-2 Pathogenic Samples Preparation
- Access controlled facility (CL2) for the use of hazard group 2 pathogens
- Class 2 microbiological safety cabinet
- PHCBI multi-gas incubator (MCO-50M-PE IncuSafe)
- Temporary storage space (4°C and liquid nitrogen)
- Recirculating fume hood
