Kultchitzky’s Double Embedding Method
Kultchitzky also proposed a technique which involved regular celloidin processing, hardening with chloroform vapor and infiltrating with paraffin wax. It is noted as being recommended for embryos and hard insects (flees, ticks).
Procedure
- Prepare a 1% solution of celloidin in methyl benzoate. This should have the consistency of honey.
- Thoroughly fix the tissue and trim to an appropriate size no more than 3 mm thick.
- Thoroughly dehydrate the tissue to the absolute ethanol stage.
- Place in an equal parts methyl benzoate-ethanol mixture for a few hours.
- Infiltrate with celloidin in methyl benzoate overnight.
- Drain excess celloidin from the tissue and place tissue into xylene, 3 changes, an hour in each.
- Infiltrate with paraffin wax at 60°C for 3 changes of 2 hours each with intermittent agitation.
- Cast the block as for the paraffin infiltration technique.
- Section, float out, stain and coverslip as for a paraffin block.
Safety Note
Prior to handling any chemical, consult the Safety Data Sheet (SDS) for proper handling and safety precautions.
References
- Gatenby, J. B. and Beams, H. W.,(1950)
The Microtomist’s Vade-Mecum Ed. 11, pp. 100-108, (para. 187-203).
J. & A Churchill Ltd., London, UK.
