Tissue Culture & Sample Preparation Facility

Tissue Culture Facility

The WUCCI tissue culture facility features state-of-the-art equipment for all necessary BSL-2 culture conditions. The facility is enclosed in a dedicated space within the WUCCI complex and is equipped with two 4’ Class II, Type A2 bio safety cabinets and four HeraCell VIOS copper lined, self-sterilizing incubators (two CO2 and two tri-gas for hypoxic conditions). The facility also houses a Sorvall X1R refrigerated, swinging bucket centrifuge, a 12 L dry bath, a fridge/freezer, and wet-prep bench space.

Sample Preparation Facility

Thin Sectioning

1. Leica UC7 Ultramicrotome

The thin sectioning room houses two UC7 ultramicrotomes for routine and ultrathin EM sectioning. These units both have touchscreen control, Multi-LED illumination, eucentric movement for the microscope, and automated routines for trimming and sectioning. In addition, each unit features a digital camera coupled to a 50” LED TV for ergonomic and teaching purposes.

For Tokuyasu or cryo-sectioning, the center also has a Leica FC7 cryo stage attachment for the UC7. This unit boasts three temperate zones for precise sectioning control, anti-contamination airflow, and a CRION antistatic device with discharge mode.

2. Leica KMR3 Glass Knife Maker

The Leica EM KMR3 knifemaker is a complete tool-free unit that scores, clamps, and breaks glass strips into knives for ultramicrotomy. Available glass strips are 6.4mm thick.

Sample Preparation

1. Leica EM ICE High Pressure Freezer

The EM ICE is a unique workstation that combines both high-pressure freezing and light stimulation with millisecond precision. Unlike previous approaches, in this case no alcohol or cryoprotectants are required for operation, and an automated loading system allows for one-second fresh-to-frozen cycles. In addition, the system allows for freezing specimens up to 5mm in diameter, and accommodates a variety of sample holders including sapphire disks (3 mm and 6 mm), freeze fracture holders, and membrane carriers.

Light stimulation is achieved via easily exchangeable LED light modules. Currently, the system has blue (450 λ), green (523 λ) and amber (597 λ) modules. Light stimulation is achieved by easy to program routines through the system software and can be customized for any duration or number of pulses. After stimulation, the sample is instantly frozen within one millisecond, precisely capturing any dynamic process.

2. Leica AFS2 Freeze Substitution Unit

The AFS2 is a fully automated freeze substitution system for processing samples after they have been high pressure frozen. It also includes the EM FSP automatic robotic processor with integrated UV LEDs for user-free operation.

3. Leica CPD300 Critical Point Drier

The EM CPD300 is a fully automated critical point drier for the gentle drying of samples for SEM analysis. It is compatible with samples that have been dehydrated in ethanol, methanol, acetone, and isopropyl alcohol. This unit can accommodate small to medium (~75 x 40mm) sized samples, and has dedicated holders for drying multiple coverslips. Runs can be fully customized with number of cycles and fluid exchange speeds to accommodate a large variety of sample types.

4. Leica ACE600 Sputter / Carbon Coater

The ACE600 coater is a high-resolution turbo-pumped combination sputter coater, carbon thread evaporator, and glow discharge device. It features a motorized, planetary drive stage with quartz crystal thickness monitor. Foil targets for sputtering include: Iridium, Platinum, and Gold.