Abstract
Continuous observation of embryonic growth can improve understanding of the early developmental events and allow us to use parametric statistical analyses with time as a parameter. A cinematographic study such as that reported here utilizes time-lapse video recording. Previously published methods for time-lapse video recording have involved building an incubator around a microscope, a process that is both expensive and laborious. Here we present a simplified method for time-lapse video recording of early bovine embryo development. The embryos were cultured during a 24-hour period in a standard pregassed tissue culture bottle, which was darkened and placed on the heating stage of an inverted microscope for recording through a red filter. The control embryos were cultured in a conventional CO2 incubator. After 10 replicates we could not find a statistically significant difference between the cell numbers of these two treatments (P=0.95), suggesting that the culture setup is appropriate for continuous observation of early cleavage of the cattle embryo.
Highlights
Improvements in the in vitro production of embryos depend on well-designed embryo culture experiments
Continuous observation facilitates more versatile use bf parametric statistical analyses, as time can be used as a parameter
The setup has usually been based on a C02 incubator built around a microscope
Summary
Improvements in the in vitro production of embryos depend on well-designed embryo culture experiments. Continuous observation of embryo culture produces considerably more information than the end point approach. Time-lapse video recording of embryos week of culture. Continuous monitoring of embryo development makes use of time-lapse video recording. We present here a simple and inexpensive culture system designed for time-lapse video recording of early bovine embryo development. We compare the final cell numbers at the end of 24-h culture between the time-lapse video recording and regular C02 incubator environments. Using red light (> 620 nm) illumination caused by a filter placed over the window at the top of the flask, we recorded the culture period at 10 x magnification with a Hamamatsu C4200 CCD microscope camera and a time-lapse video recorder (Fig. 2). At 44 hpi both embryo cultures were terminated and the cleavage rates and the cell numbers per cleaved embryo were calculated in both groups
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