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Maslakova lab advice on toxic and non-toxic materials for embryology (adapted from R. Strathmann at FHL) Areas in which we work with embryos and larvae (“E areas”) need to be kept as free as possible of agents that threaten long-term culture viability. A good rule of thumb is that if you wouldn't rub it in your eyes, don't bring it into an E area. Remember that if you handle toxic materials in the next lab over, you might bring it in on your hands. The three most common problems: • • • Detergents – rinse hands carefully and keep detergent bottles away from E sinks and E glassware. Hand lotion, sunscreen lotion, etc. – if you must use lotions, be careful to keep fingers out of cultures. Fixatives – keep fixatives as far as practical from all cultures. As much as possible, do all fixations and handling of fixed samples in a non-E area (e.g. in another lab, or in the fume hood). Avoid bringing pipettes or vials or glassware used for fixatives into an E area. For cultures in standing or recirculating water the following common laboratory materials are Probably toxic Usually non-toxic latex tubing, other rubber tubing, rubber stoppers polyethylene and tygon tubing (after rinse) any metal object or heavy metals in solution acrylic plastic, glass, polyethylene paper towels, possibly PVC (well rinsed to remove catalysts or solvents) lotions and creams, anything else that might leave oily or greasy residues hot-melt glue and clear silicone glue (after solvent evaporates) detergents paraffin and parafilm fixatives, of course nylon netting, Nitex ink bag filters, glass filters, membrane filters most adhesives, including tape of all kinds most paper filters after rinse Corning high-vacuum grease, Vaseline mineral oil, halocarbon oil E-ware: Glassware and other materials for cultures should be labelled "E" and should not be contaminated. If glassware is contaminated, remove the "E" and place it with non-E stock. E glassware should be exposed only to seawater, tap water, distilled or RO water, and, for cleaning, HCl and ethanol (which leave no residues) Plastic-fiber scrubbers (without detergent, sold for household use) or standard lab brushes (labeled only for E use) can be used to remove organic material from E-ware without solvents. New objects should be rinsed before use to remove residues from the factory or storage. Other points: • Many micro-pore membrane filters include detergents that increase wetability. When in doubt, discard the first water through such filters. • Some larvae are sensitive to components of algal enrichment media. In general medium should be removed from algae used for food by centrifugation and resuspension of algal cells in sea water. Removal of algal medium is not essential for routine feeding of many kinds of larvae, however. • Make sure clothes-pins used to label cultures are not shedding metal fragments.