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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:
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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.
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Specific
procedures for working with crystals at CHESS are given below: |
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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.
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Virus crystals will be
usually be kept in the F-1 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 or if they are being used
at the A-1 or F-2 stations). The cognizant Safety Officer, or his/her
designate, is to verify the nature and amount of viral materials brought
by the user.
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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.
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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.
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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.
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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, or in the F-1 Biosafety Cabinet. Flammable
materials such as propane are not to be used in the Biosafety Cabinet.
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Loaded goniometer heads will
be carried from the storage and mounting area into the hutch enclosed in
sealed, non-breakable containers. If such a container is dropped, any
spill of viral material will be contained within it. 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.
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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.
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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.
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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. |
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Guidelines
last revised February 2001. |
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