Title: A Template for Insect Cryopreservation Authors
Submitted to: Book Chapter
Publication Type: Book / Chapter
Publication Acceptance Date: September 17, 2009
Publication Date: January 1, 2010
Repository URL: http://ddr.nal.usda.gov/bitstream/10113/41339/1/IND44364787.pdf
Citation: Leopold, R.A., Rinehart, J.P. 2010. A Template for Insect Cryopreservation. In: Denlinger, D.L., Lee, R.E. Jr., editors. Low Temperature Biology of Insects. Cambridge University Press. p. 323-341. Technical Abstract: This article is intended to update the reader on the progress made on insect embryo cryopreservation in the past 20 years and gives information for developing a protocol for cryopreserving insects by using a 2001 study as a template. The study used for the template is the cryopreservation of the Old World Screwworm, Cochliomyia hominivorax. To supplement the information on protocol development, a generalized theoretical basis for formulating each step in the cryopreservation procedure is also included in this chapter. The topics included in the theoretical basis section are: prevention of intracellular ice formation, embryo permeabilization, slow vs. rapid cooling, cryoprotective agents, osmotic equilibrium, and pre- and post-treatment handling of embryos. Following this section, is instruction on optimization of certain components of the cryopreservation protocol where modifications can help improve the overall yield of viable insects following storage in liquid nitrogen. Protocol optimization is often necessary because there are a number of species-specific elements that require individual customization to gain peak production of viable adults. Complementing the instructions on protocol development are photomicrographs showing screwworm embryos during treatment prior to storage in liquid nitrogen and tables relating to process of cryopreserving this insect. Thus, the primary goal of this article is to inform, instruct and generate interest on the relative ease of cryopreserving insect germplasm for use in research, control and other issues associated with human health and welfare.