A simple method for counting nuclei in the preimplantation mouse embryo

K. M. Ebert, Robert E Hammer, V. E. Papaioannou

Research output: Contribution to journalArticle

50 Scopus citations

Abstract

An easy and rapid method of counting the number of cells in the preimplantation mouse embryo is described. The procedure increases the speed with which large numbers of embryos can be processed using a simple squash technique. Cell numbers are determined by exposing the embryos to the fluorescent DNA-binding dye, Hoechst 33258, removing the zona pellucida and simply squashing the embryo and counting the number of fluorescent nuclei. An increase in fluorescent intensity and maintenance of nuclear conformation of the squashed preparations are greatly improved by the use of the non-ionic detergent Triton X-100. Viability of dye-treated fertilized one-cell and blastocyst stage embryos is maintained at least up to day 13 of pregnancy following transfer of the embryos to the uteri of pseudopregnant recipients. Additional uses for this staining technique are discussed.

Original languageEnglish (US)
Pages (from-to)1207-1209
Number of pages3
JournalExperientia
Volume41
Issue number9
DOIs
Publication statusPublished - Sep 1985

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Keywords

  • blastocyst
  • Hoechst 33258
  • Morula
  • nuclei
  • vital stain

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

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