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.

Squashy Eggs: Technical Note

J. Carrel

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.