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A rapid procedure for visualising the inner cell mass and trophectoderm nuclei of mouse blastocysts in situ using polynucleotide‐specific fluorochromes

A rapid procedure for visualising the inner cell mass and trophectoderm nuclei of mouse... A rapid procedure has been devised to count the numbers of outer trophectoderm (TE) and inner cell mass (ICM) cells of mouse blastocysts by differentially labelling their nuclei in situ with polynucleotide‐specific fluorochromes. The TE nuclei were labelled with propidium iodide (PI) by permeabilising the cells using selective antibody‐mediated complement lysis (Solter and Knowles, '75). The blastocysts were then fixed in ethanol and the ICM nuclei labelled with bisbenzimide. These two fluorochromes have widely different fluorescent spectra. Thus, by using fluorescence microscopy with appropriate filter combinations, the PI‐labelled TE nuclei appeared pink or red; the bisbenzimide‐labelled ICM nuclei, blue or unlabelled. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png The Journal of Experimental Zoology Part A: Ecological and Integrative Physiology Wiley

A rapid procedure for visualising the inner cell mass and trophectoderm nuclei of mouse blastocysts in situ using polynucleotide‐specific fluorochromes

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References (22)

Publisher
Wiley
Copyright
Copyright © 1984 Wiley Subscription Services, Inc., A Wiley Company
ISSN
2471-5638
eISSN
2471-5646
DOI
10.1002/jez.1402310317
pmid
6209359
Publisher site
See Article on Publisher Site

Abstract

A rapid procedure has been devised to count the numbers of outer trophectoderm (TE) and inner cell mass (ICM) cells of mouse blastocysts by differentially labelling their nuclei in situ with polynucleotide‐specific fluorochromes. The TE nuclei were labelled with propidium iodide (PI) by permeabilising the cells using selective antibody‐mediated complement lysis (Solter and Knowles, '75). The blastocysts were then fixed in ethanol and the ICM nuclei labelled with bisbenzimide. These two fluorochromes have widely different fluorescent spectra. Thus, by using fluorescence microscopy with appropriate filter combinations, the PI‐labelled TE nuclei appeared pink or red; the bisbenzimide‐labelled ICM nuclei, blue or unlabelled.

Journal

The Journal of Experimental Zoology Part A: Ecological and Integrative Physiology – Wiley

Published: Sep 1, 1984

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