Worm Breeder's Gazette 1(2): 8b
These abstracts should not be cited in bibliographies. Material contained herein should be treated as personal communication and should be cited as such only with the consent of the author.
We have investigated the use of hypochlorite and hydrophobic agents to permeabilize nematode eggs without altering viability. Nucleopore membrane filters, 8 m in pore size, are useful because they soak up little fluid, retain eggs well by adhesion, and are almost transparent. By washing eggs gently for 2 minutes with 2% hypochlorite, we have made them 100% susceptible to 1% SDS while retaining 100% viability. A hydrophobic barrier probably remains as the eggs still remain insensitive to glutaraldehyde. More vigorous washing or further treatment increases susceptibility to glutaraldehyde but also decreases the viability of the eggs. Permeabilized eggs stain with neutral red iodide (1 mg/100 ml). We have found mutants whose eggs readily take up neutral red but the usefulness of the mutants is impaired by the low viability of the eggs. A relatively well-synchronized (~l hour) but significantly inviable ( ~50%) population of eggs can be obtained as follows. Synchronize larvae using the egg wash method (Russell and Hirsh labs). Just as the larvae reach sexual maturity and are producing the first eggs, dissolve worms in 2% hypochlorite (3 to 4 minutes required) and gently wash eggs on membrane filter.