Dear Arthur,

Thank you for your interest, and my apologies for taking a while to respond.

For fixing head lice at diffrent stages of oviposition, I used liquid "canned
air" to stop them in their tracks, then tipped in an excess of hot alcoholic
Bouin (or Dubosq Brasil as we entomologists called it).  They were kept at 60
degrees for 1 hour, then transferred to alcohol for dehydration (Automatic
embedder, 45 minute changes at 80%, 96% then 100% ethanol), clearing in
toluene (2 x 45 min) and embedding in soft (mp 60 degrees) paraffin wax (2 x
45 min).  This worked better than formaldehyde, gluteraldehyde, Susa, regular
Bouin, or Carnoy for histological preservation.  Fixing punctured insects or
removed abdomens gave no benefit in preservation.  I understand Picric acid
fixatives preserve antigenicity at least as well as formalin.

For dissecting out the reproductive system of head lice, I sprinkled them on
a wax dish and used a soldering iron to melt in front of their head so they
are trapped thorax deep in wax.  By slicing off the tip of the abdomen with a
mounted razor, all the innards can be squeezed out with fine angled forceps
and fixed in formaldehyde.  This got my dissection time down to a few seconds
each and allowed me to do SEM of the internal organs.

For making transparent whole mounts of fleas, the insects can be collected in
30% alcohol.  I can't lay my hands on the exact protocol, but know it came
from one of Mirriam Rothschild's old papers.  The short cut was to put one
pellet of KOH in an eppendorf with a ml of water.  When dissolved, I added
the fleas and kept it in a 60 degree incubator for I think 4 hours.  The
longer you leave them, the lighter they get.  Embedding was done through
alcohol, then oil of clove (try Xylene), then DPX.

Good luck

David Carter
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