Michael L. Reed’s research while affiliated with Reproductive Medicine Institute and other places

In clinical IVF, the incubator is a surrogate oviduct and uterus. During this stage, the embryo normally travels through a dynamic environment from the oviduct to the uterus. Temperature, pH, osmolality, and many other environmental factors change during its journey. Current incubators and culture media are fairly static. The incubator must often provide an environment that is a compromise—suitable for sperm, oocytes, and the various stages of preimplantation embryo development. This system should not just address ideal growth but should also allow for ideal expression of embryonic genes. It should provide for the necessary chemicals, growth factors, proteins, osmolality, temperature, and pH. Almost all current culture media have been developed for mice gametes and embryos and then applied to humans. In addition, we often are not aware of the ideal conditions for human gametes and embryos.

Purpose To determine liquid nitrogen evaporation rates of intact liquid nitrogen storage tanks and tanks with their vacuum removed. Methods Donated storage tank performance (LN2 evaporation) was evaluated before and after induced vacuum failure. Vacuum of each tank was removed by drilling through the vacuum port. Temperature probes were placed 2 in. below the bottom of the styrofoam cap/plug, and tanks were weighed every 3 h. Evaporation rate and time from failure to the critical temperature was determined. Result Storage tanks with failed vacuum have a much higher evaporation rate than those with intact vacuum; evaporation rates increased dramatically within 3 to 6 h in the smaller tanks, and time to complete depletion varied according to starting LN2 volume. Tanks with storage racks/specimens may have altered evaporation profiles compared to tanks without. Locating temperature probes 2 in. below the styrofoam cap/plug suggests that for most applications, alarms would sound approximately 1 h prior to reaching the critical warming temperature, approximately − 130 °C. External signs of vacuum loss were dramatic: vapor, frost, and audible movement of air. Conclusion For the first time, we have data on how liquid nitrogen storage tanks behave when their vacuum is removed. These findings are conservative; each lab must consider starting volume, tank size/capacity, function (storage or shipping), age, and pre-existing evaporation behavior in order to develop an emergency response to critical tank failure. Times to complete failure/evaporation and critical warming temperature after vacuum loss are different; these data should be considered when evaluating tank alarm systems.

Maintaining consistent and reliably high success rates is a daily challenge for every IVF laboratory. This step-by-step guide is an essential aid in navigating the complex maze of physical, chemical, biological, and logistic parameters that underpin successful gamete and embryo culture: temperature, pH, osmolality, gas supplies, air quality, light exposure, infections, managing supplies, personnel, as well as overall quality control. Numerous real-life troubleshooting case reports are presented, identifying all aspects necessary for troubleshooting. Process maps and flow charts accompanying each chapter offer a logical and systematic approach to problem solving in the laboratory. This is an essential resource for scientists in assisted reproductive technology and specialists in reproductive biology and medicine, helping IVF clinics to achieve the dream of every infertile couple: the birth of a healthy child.

In oviparous animals, the egg hatches outside of the body and is exposed to light; in some cases throughout the development of the fetus. In mammals, fertilization and the growth of embryos in vivo occurs in the dark but in human IVF, these embryos are exposed to variable light sources and intensities. Light can affect embryonic development in some species via either a direct toxic effect on the embryo, or indirectly via photo-oxidation of components in the media or oil. Although data regarding the effect of light on human embryos is lacking, it is prudent to take appropriate steps to minimize the potential harmful effects of both ambient and microscopic light on embryos.