All new users must read User Orientation and Safety Training Guide, and score 100% on the short User Online exam, before beginning any work at CHESS.
Prepare for your visit
General User Information
The CHESS New User Guide is a step-by-step guide to help you to apply for beamtime, and once accepted, how to get ready for your experiment, what to do when you get here and how to report when you leave.
User Orientation and Safety Training
As a prerequisite for work at CHESS it must be established whether or not Hazardous Materials are involved. If such materials are to be brought to CHESS, certain information must be provided and internal procedures followed before actual beam-time is granted. These procedures involve two parts, (1) those that must be completed well in advance of arrival at CHESS and (2) those to be carried out when the hazardous material actually arrives at CHESS. Upon submission of your proposal, any hazardous or potentially hazardous, explosive, toxic, radioactive materials, or radiation producing sources, must be declared and submitted in writing for review by the CHESS Safety Committee. The following information is required for EACH MATERIAL.
- Name of material (one form for each material)
- MSDS Sheet (MSDS CHESS inventory)
- Description of substance (powder, liquid, solution etc.)
- Amount (several pounds, several micrograms)
- Means of containment (sealed in glass capillaries, flasks, encapsulated)
- Toxicity routes (ingestion, subcutaneous, absorption, inhalation)
- Procedures in case of emergency (clean-up, evacuation, emergency first-aid).
Especially useful to us are the safety precautions your home institution requires or suggests in the proper handling of the material and what actions should be taken in the event of an emergency.
All proposals that are declared hazardous will be assigned a Safety Officer (a member of the CHESS Safety Committee) to check-in the group when they arrive at CHESS and check-out the group when they leave. The Safety Officer will ensure that all equipment is safe to use, will go over detailed safety information related to the experiment, and will arrange disposal of any hazardous waste. Contact the CHESS Safety Committee at firstname.lastname@example.org with any questions regarding hazardous materials and safety concerns.
Users are encouraged to bring the smallest amounts possible of hazardous materials to CHESS.
Please do not bring the hazardous materials into the building before you meet with a member of our Safety Committee. The operator on duty will, upon your checking in, put you in contact with this person. In certain instances (such as with gas cylinders) a preliminary inspection will be required before entry of the material into the building is allowed.
Brief guide: hazardous vs. non-hazardous
An experiment is non-hazardous if:
- The protein or other material being studied is not toxic (or infectious) for humans.
- No heavy atom compounds are used (intrinsic metals such as the Fe in hemoglobin are not considered as heavy atoms, nor are metal foil reference samples).
- Heavy atom compounds are present only in pre-derivatized crystals.
- The crystals may be mounted in capillaries, frozen, or contained in solutions free of the heavy atom compound.
- As long as you stay within this category, CHESS does not need to be informed if you bring samples other than those listed on the proposal.
An experiment may be hazardous if:
- Heavy atom compounds are present outside of crystals. In this case, you must supply CHESS with a Heavy Atom Compounds Declaration, listing the type and amount of each compound. The Safety Committee will then classify the experiment as hazardous or not, using the following guidelines:
- For solutions:
The experiment is considered non-hazardous if the total quantity of each heavy atom compound is no more than 1/50 the LD-50 for a 50 kg. animal or 0.01 mM if the LD-50 is not available. LD-50's, when known, are given on Material Safety Data Sheets. If multiple values are given the most appropriate one (in terms of method of administration and similarity of test animal to humans) is used. If the total quantity is above these values but below 1/2 the LD-50 for a 50 kg. animal, or 0.25 mM if the LD-50 is not available, the experiment is hazardous but may be carried out at CHESS. Quantities greater than 1/2 the LD-50 or 0.25 mM may not be brought to CHESS.
For dry material:
Reference samples for MAD experiments are non-hazardous if they are well-sealed, e.g. between two layers of Kapton tape, before being brought to CHESS. For loose material, the same limits on total quantity as for solutions are used.
- For solutions:
An experiment is hazardous if:
- The protein or other material is toxic or infectious to humans. In this case, you must supply CHESS with a Hazardous Materials Declaration, and Viral Materials Declaration, if appropriate. Extra time will be needed to review this documentation.
If classified as hazardous, extra time will be needed for the proposal to be processed.
If in doubt, contact Marian Szebenyi, (607) 255-5717, for clarification.
Procedures to be used with BL-2 viruses at CHESS
Procedures to be used with BL-2 viruses
In general, NIH/CDC guidelines for dealing with BL-2 material should be followed. Relevant practices for areas in which viruses will be handled include:
No eating, drinking, smoking, applying contacts or cosmetics; no pipetting by mouth. Wear appropriate personal protective equipment (PPE) - at least gloves and lab coat. Wash hands after removing gloves or handling biohazardous material. Restrict personnel access while work with biohazards is in progress. Decontaminate work surfaces at least once a day.
Specific procedures for working with crystals at CHESS are given below:
- Crystals of the virus will be brought to CHESS in a double-sealed, appropriately labeled container, as required by the DOT for transport of infectious materials. Crystallization trays or sealed capillaries, containing unfrozen crystals, will be surrounded with absorbent material and sealed within a sturdy outer container. Frozen crystals will be contained in vials enclosed in a Dewar or dry shipper; the container will be secured so that it cannot tip over.
- Virus crystals will be kept in the station. However, they may also be stored in another room, such as the cold room or the chemistry lab, if necessary (e.g. if crystals must be mounted in the cold). The cognizant Safety Officer, or his/her designate, is to verify the nature and amount of viral materials brought by the user.
- Biohazard warning signs will be placed on the doors of all rooms in which crystals will be stored or used. Access to areas in which crystals will be handled will be restricted to the investigators, the CHESS and MacCHESS operators, and members of the CHESS and LNS Safety Committees. If other personnel require access to such an area, the crystals shall be first contained in a suitable box, except for a crystal actually mounted on the oscillation camera.
- Table and bench tops in the storage and mounting area will be covered with plastic-backed absorbent paper, which is soft enough so that any dropped capillary or crystal will not bounce off, and absorbent enough to soak up any spills. When crystals are actually being manipulated, it is recommended to cover the immediate area with absorbent paper wetted with disinfectant, so that any dropped crystal will be inactivated.
- In the hutch, any surfaces on which a capillary or crystal might fall will be covered with plastic-backed absorbent paper. If frozen crystals are being used, any equipment which could be contaminated in case of failure of the cryosystem (e.g. the rotation stage) should be protected by a plastic shield which can be removed and decontaminated. Freezing of crystals directly into the cold stream is permitted, provided coverage of surfaces and equipment is adequate to catch any viral material which could be dropped in the mounting process. A germicidal lamp will be available in the hutch, in case decontamination is needed.
- Mounting of crystals into capillaries, or freezing of crystals into liquid nitrogen (or other substance such as propane), will be performed in the room where the crystals are stored.
- Crystals frozen in the storage and mounting area will be transported to the hutch in a container of liquid nitrogen which is enclosed in an outer container to catch any spills.
- Because of the containment procedures given above, any spill (e.g. a dropped crystal or a broken capillary) should be limited to an absorbent paper-covered surface or the interior of a container. The users will clean up the spill, using bleach or other suitable disinfectant to decontaminate any places (e.g. absorbent paper or the inside of the outer container) where viral material might have landed. Broken capillaries are to be placed in a "sharps" container for later disposal. In the unlikely case of a spill which involves unprotected surfaces or equipment, or in which an aerosol may have been produced, the cognizant Safety Officer will be consulted to determine appropriate decontamination procedures. A reminder of procedures to follow in case of spills will be posted in the crystal mounting and storage area, and in the hutch.
- Used capillaries are to be placed in a "sharps" container. Used frozen crystals are to be either disposed of in a biohazard bag (use disinfectant solution to wash a crystal out of a cryoloop into a small container and put the container in the bag, if the loop is to be salvaged) or stored in a Dewar, still frozen, for possible later use.
- At the completion of their experiments, the users will roll up the absorbent paper and dispose of it in a biohazard bag. All surfaces which could have been contacted by virus will be washed with a suitable disinfectant, e.g. bleach or alcohol. The materials used for this cleanup, e.g. paper towels, will also be disposed of in the biohazard bag. Biohazard bags and sharps containers of used capillaries will be placed in a biohazard container (provided by CHESS) for later pickup by Cornell EH&S. The cognizant Safety Officer, or designate, is to verify that no biohazardous materials have been left at CHESS, except in the biohazard container.