This guide provides the necessary steps to prepare samples at room temperature for submission to the cryo-microscopy service.

• Protection: Ensure that all samples are packaged securely to prevent contamination, spills, or damage during transit.
• Sealing: Use leak-proof containers or sealed tubes to avoid any leakage.

• Tube Labels: Clearly label each tube with the sample name, concentration, and any other relevant details.
• Avoid Mislabeling: Make sure the labels are securely attached and legible to prevent misidentification.

• Buffer Addition: Ensure that all necessary buffers are included with the samples if any dilutions or additional preparation steps are required.
• Concentration: Indicate the concentration of each buffer and ensure that they are compatible with cryo-microscopy requirements.

• Risk Identification: Clearly define and label any samples containing chemical or biological hazards.
• Hazard Labels: Use appropriate hazard symbols and indicate any special handling requirements on the containers and accompanying documentation.

• Tube Labels: As with room temperature samples, clearly label each tube with the sample name, concentration, and any additional relevant details.
• Hazard Labels: If the samples contain chemical or biological hazards, add appropriate labeling and hazard symbols.
• Documentation: Include documentation specifying the temperature requirements (4°C or -20°C), and any handling instructions.

• Packaging: Use a strong, insulated container (such as a styrofoam box) that can withstand dry ice temperatures.
• Dry Ice Quantity: Ensure there is enough dry ice to maintain -20°C for the full duration of shipping.
• Labeling: Mark the container with “Keep at -20°C” and include any relevant safety warnings about dry ice.
• Ventilation: Ensure the container is ventilated to allow CO₂ gas to escape, preventing pressure buildup.

• Packaging: Place samples in a sealed container to prevent moisture ingress and label them as "Keep at 4°C".
• Cooling Method: Use an adequate amount of ice to keep the samples at a stable temperature of 4°C during transit.
• Insulation: Pack the samples in a styrofoam container or insulated box to maintain a stable temperature.
• Duration: For longer shipping times, check the amount of ice required to ensure temperature consistency upon arrival.

For samples prepared on vitrified grids, please follow these instructions carefully. Vitrified samples must be preserved at cryogenic temperatures using a dry shipper to prevent thawing or damage.

• Condition Documentation: Clearly document the conditions for each grid, including:

  • Grid Material: Specify the type of material (e.g., copper, gold).
  • Hole Size: Indicate the hole size (e.g., 1.2 µm, 2.0 µm).
  • Other Conditions: Note any additional parameters, such as specific coating or sample concentration.

• Grid Box Labeling: Use a GridBox to organize and secure each grid during shipment.

  • Grid Placement Diagram: Draw a simple diagram showing the layout and positioning of each grid within the GridBox.
  • Labeling: Clearly label each slot in the GridBox to match the corresponding grid's documented conditions.

• Dry Shipper Preparation: Ensure that the dry shipper is pre-cooled with liquid nitrogen and emptied of any remaining liquid before packing the grids to avoid contamination.

• GridBox Placement: Secure the GridBox within the dry shipper, ensuring it is stable and will not shift during transit, pucks or tubes.
• Ventilation: Make sure the dry shipper allows ventilation for gas release, preventing pressure buildup on the tube.

• Container Labeling: Mark the dry shipper as Cryogenic Material and include appropriate hazard labels for safe handling.
• Grid Details: Include a printed copy of the grid conditions, including the layout diagram and any specific handling instructions.
• Temperature Requirements: Clearly state that samples must remain at cryogenic temperatures to maintain integrity.

• ✅ Include Buffer for dilutions.
• ✅ Pack 4°C samples with sufficient ice and insulated containers.
• ✅ Use ample dry ice for -20°C samples and ventilate the container.
• ✅ Label tubes and containers clearly with temperature and hazard information.
• ✅ Include documentation with storage and handling requirements.
• ✅ Document grid conditions: material, hole size, and any other parameters.
• ✅ Draw a layout diagram for the GridBox and label each grid position.
• ✅ Prepare the dry shipper with adequate ventilation and secure the GridBox inside.
• ✅ Label the container and include documentation to return the container (dry shipper).

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C/ Darwin nº 3
Centro Nacional de Biotecnología
28049 Cantoblanco
Madrid

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For any further questions, please contact the cryo-microscopy service team. [email protected]