{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,2]],"date-time":"2026-06-02T23:28:22Z","timestamp":1780442902042,"version":"3.54.1"},"reference-count":32,"publisher":"MDPI AG","issue":"10","license":[{"start":{"date-parts":[[2022,10,20]],"date-time":"2022-10-20T00:00:00Z","timestamp":1666224000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"U.S. Department of Agriculture, Agricultural Research Service","award":["3060-21000-041-00D"],"award-info":[{"award-number":["3060-21000-041-00D"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Insects"],"abstract":"<jats:p>Embryos of the sunflower moth, Homoeosoma electellum (Hulst), were cryopreserved after modification to the method that was previously described for Pectinophora gossipiella. The workflow to develop the protocol consisted of methods to weaken the embryonic chorion followed by the application of various methods to disrupt the sub-chorionic wax layer. These steps were necessary to render the embryos permeable to water and cryoprotectants. Initially, the embryos were incubated at 21\u00b0 and 24 \u00b0C, and the development of the double pigment spots\/eyespot and eclosion were tracked every two hours. The embryos at 24 \u00b0C showed eyespots as early as 30 h, while in the case of the embryos that were incubated at 21 \u00b0C, there was a developmental delay of approximately 20 h. The embryos at 24 \u00b0C showed peak eclosion between 55 and 70 h, and the embryos at 21 \u00b0C eclosed between 80 and 100 h of development. Estimating this range is crucial for the purposes of stage selection and treatment initiation for cryopreservation protocol development for the embryos. The control hatch percentage at either developmental temperature was &gt;90%, and the sodium hypochloride, 2-propanol and alkane-based treatments reduced the embryo hatchability to &lt;10%. Hence, a modified surfactant\u2014hypochlorite mixture\u2014was used to destabilize the chorion and solubilize the hydrophobic lipid layers. Water permeability assessments using the dye-uptake method show that polysorbate 80 in combination with sodium hypochlorite alone is capable of permeabilizing the embryo as efficiently as sequential hypochlorite\u2014alkane treatments, but with significantly higher hatch rates. A vitrification medium consisting of ethane diol and trehalose was used to dehydrate and load the embryos with the cryoprotective agent. The median hatch rates after vitrification were 10%, and maximum was 23%.<\/jats:p>","DOI":"10.3390\/insects13100959","type":"journal-article","created":{"date-parts":[[2022,10,20]],"date-time":"2022-10-20T22:49:46Z","timestamp":1666306186000},"page":"959","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Vitrification of Lepidopteran Embryos\u2014A Simple Protocol to Cryopreserve the Embryos of the Sunflower Moth, Homoeosoma electellum"],"prefix":"10.3390","volume":"13","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-1587-2891","authenticated-orcid":false,"given":"Arun","family":"Rajamohan","sequence":"first","affiliation":[{"name":"Insect Biochemistry and Genetics Research Unit, USDA-ARS-Edward T. Schafer Agricultural Research Center, 1616 Albrecht Blvd., Fargo, ND 58102, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6165-7319","authenticated-orcid":false,"given":"Jarrad R.","family":"Prasifka","sequence":"additional","affiliation":[{"name":"Sunflower and Plant Biology Research Unit, USDA-ARS-Edward T. Schafer Agricultural Research Center, 1616 Albrecht Blvd., Fargo, ND 58102, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Joseph P.","family":"Rinehart","sequence":"additional","affiliation":[{"name":"Insect Biochemistry and Genetics Research Unit, USDA-ARS-Edward T. Schafer Agricultural Research Center, 1616 Albrecht Blvd., Fargo, ND 58102, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,10,20]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Lee, R.E., and Denlinger, D.L. (1991). 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