1. 2.

Tools used on living material should be separate from those used with fixative because fixative is almost impossible to remove completely from any tool. Fixative, even at a very small amount, can severely damage living material, including those from embryos. 3. Waterstone is used instead of oilstone in order to avoid oil contamination on tools. Soft stone should be used for the first honing, and a harder stone should be used for final polishing. The stone should always be kept wet with water when in use to keep metal particles from clogging the stone's pores. The stone should be washed with water after use in order to remove metal residues. The whole surface of the stone should be used to avoid wearing it unevenly. The stone should be stored dry in a soft cloth bag. 4. Workbench -If available, a culture hood should be used: one painted with high-gloss enamel, and using a work light and a germicidal UV light. -The hood should be located in an area free from drafts from doors, windows, and other equipment (Gibco, -An opaque curtain or coated bench paper should cover the hood when the UV light is on -Leave the UV light on for one hour to overnight before work -Never work under the hood with the UV light on -Never leave cultures in the hood with the UV light on -Workspace on the lab table should be wiped down with 70% alcohol -The work surface should only contain items to be used for the specific procedure (Gibco -Stage and knobs of dissecting microscope should be wiped down with 70% alcohol -Instrument tray and all items should be used should be wiped down with 70% alcohol -Do not talk, sing, or whistle when performing sterile procedures (Gibco -Perform experiments as rapidly as possible to minimize the possibility of contamination (Gibco Dissecting Tools -Tools should be sterilized and resterilized after each use by soaking for 10 minutes in 70% alcohol, placing in bouling water, or autoclaving -For soaking, a lidded jar with cotton at the bottom should be filled with 70% alcohol -Do not leave alcohol on the instrument when it comes into contact with culture tissue -Everytime an instrument is used, dip it back in the alcohol and return into the instrument tray Glassware

-All glassware must be cleaned carefully using detergent that is safe for culture use -Glassware should be rinsed and re-rinsed carefully in distilled water and air-dried -Glassware should be wrapped in aluminum foil after drying, folding over each seam twice -Glassware without stoppers or tops can be autoclaved in dry heat. Otherwise, it must be autoclaved in moist heat. -Stoppers should be covered in foil -Plastic tops should be screwed on loosely -Presterilized plastic dishes can be used Fluids -Never uncover sterile flasks, bottles, petri dishes, etc, until they are ready to be used, and never leave them open to the environment (Gibco -Avoid pouring reagents and media directly from flasks or bottles (Gibco -Water can be boiled for sterilization -When autoclaving, bottles should be filled less than 3/4 full, and tops should be loose -Do not autoclave or boil physiological saline solutions. Instead, use a filter with a porosite of 0.22 micrometers or less. Use a sterile bottle when filtering to store the fluid in.