UROP Presentation: Optimization of Microinjection Procedures for Cryopreservation of Zebrafish Embryo

Abstract

The goal of my project was to determine optimal needle parameters for microinjection of zebrafish embryos and to use those specified needle parameters for the injection of three different cryoprotectants. The first phase of the project is meant to determine the needle parameters that cause the least amount of developmental impairment and mortality, while the second phase uses the best-performing needle for the injection of three different cryoprotectants that the Bischof lab has been developing a toxicity dataset for. Mortality one-day post-injection was determined to be significantly lower using the broken bee stinger needles than any other needle type. However, there were no significant differences between needle groups in the three-day hatch measurements as well as the five-day active swimming measurements. 99% propylene glycol (PG) and 50% sucrose unexpectedly behaved similarly to the control injected treatment of food coloring in embryo media. 99% DMSO was expected to have the highest mortality rates of all the treatments, and the data supported this; DMSO treatments led to significantly lower survival and activity rates than both control groups. The uninjected control had significantly higher survival rates than any injected group in either phase; this indicates the physical disruption of the membranes to be the most significant cause of mortality in these experiments.