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Steponkus, P.L., Meyers, S.P., Lynch, D.V., Gardner, L., Bronshteyn, V., Leibo, S.P., Rall, W.F., Pitt, R.E., Lin, T.-T. and MacIntyre, R.J. (1990) Cryopreservation of Drosophila melanogaster Embryos. Nature, 345, 170-172.
https://doi.org/10.1038/345170a0

has been cited by the following article:

• TITLE: First Estimation of Drosophila EPS Solution for Permeabilizing Lepidoptera Galleria mellonella Embryos

  AUTHORS: Muhamad Abidalla, Pio Federico Roversi

  KEYWORDS: Permeabilization, G. mellonella, Drosophila, D-Limonene, Eggshell

  JOURNAL NAME: Advances in Entomology, Vol.6 No.4, July 31, 2018

  ABSTRACT: The increased importance of the G. mellonella for wide range of scientific research and commercial sides will need to create a germplasm resource banking by cryopreservation. Impermeability is a fundamental limiting factor for the successful cryopreservation of arthropods embryos. The successful permeability of Drosophila embryo by using an embryo permeabilization solvent (EPS) solution encouraged this trial on G. mellonella embryos (stage of 24 hours Post-oviposition (h PO)). Permeability assessment with Rhodamine B and crystal violet dyes showed that G. mellonella embryos can be permeabilized by EPS of D-limonene that has 3 mol ethoxylated alcohol. The permeabilization for 30 sec exposure time was resulted 61.5% ± 5.8% survival rate, 31.7% ± 3.1% uptakes dyes and 40.5% ± 0.3% was the survival rate post loading in 12% Ethylene glycol (EG). The low viability after immersion in liquid nitrogen (LN) (0.6% ± 0.08%) is due to the dual toxicity of EPS and cryoprotectant (CPA) solutions. However, fluorescence images showed sufficient permeability that confirms the possibility to increase the permeability of G. mellonella embryos with EPS solution, and to have the opportunity to improve the viability after LN by improving procedures of loading and dehydration with various CPAs and exposure times, which decrease the toxicity effect.