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Live Imaging of Mitosis in the Developing Mouse Embryonic Cortex
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Phase-subtraction cell-counting method for live mouse embryos beyond the eight-cell stage.

William C Warger1, Judith A Newmark, Carol M Warner

  • 1Northeastern University, Department of Electrical and Computer Engineering, 440 Dana Building, 360 Huntington Avenue, Boston, Massachusetts 02115, USA. wwarger@ece.neu.edu

Journal of Biomedical Optics
|July 8, 2008
PubMed
Summary

A new, nontoxic microscopy technique accurately counts cells in mouse embryos up to the morula stage. This method could improve in vitro fertilization (IVF) success rates by better assessing embryo viability.

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Area of Science:

  • Embryology
  • Microscopy
  • In Vitro Fertilization

Background:

  • In vitro fertilization (IVF) has resulted in over 3 million births since 1978, but live birth rates remain low (34% in 2005).
  • Multiple embryo transfer, used to increase pregnancy chances, leads to high rates of multiple pregnancies (32% in 2005).
  • Current noninvasive embryo viability markers, like cell number and morphology, have limitations, especially in counting cells beyond the eight-cell stage.

Purpose of the Study:

  • To develop and validate a nontoxic method for accurately counting cells in early-stage embryos.
  • To assess the potential of cell counting as a reliable noninvasive viability marker in IVF.

Main Methods:

  • A novel phase-subtraction cell-counting method was developed by fusing differential interference contrast and optical quadrature microscopy.

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  • The technique was applied to live mouse embryos, counting cells from 8 to 26.
  • Method accuracy was verified using epifluorescence images of Hoechst-stained nuclei.

    • Main Results:

    • The phase-subtraction method accurately counted cells in mouse embryos ranging from 8 to 26 cells.
    • This nontoxic technique successfully quantified cell numbers through the morula stage.
    • The method demonstrated its potential as a noninvasive tool for embryo assessment.

    Conclusions:

    • The developed phase-subtraction cell-counting method is the first accurate, nontoxic technique for counting cells in mouse embryos up to the morula stage.
    • This method may significantly enhance the use of cell number as a critical embryo viability marker.
    • Adoption of this technique for human embryos in IVF clinics could potentially improve success rates and reduce multiple pregnancies.