Differential staining method for inner cell mass cells and trophoblastic cells of cattle blastulae

Abstract

The invention discloses a differential staining method for inner cell mass cells and trophoblastic cells of cattle blastulae. The method comprises the following steps: 1, carrying out immune combination on an anti-CDX2 monoclonal antibody which is adopted as a primary antibody and CDX2 molecules which are specifically expressed in the trophoblastic cells of the cattle blastulae through an immunostaining process; 2, the cattle blastulae which are cleaned and are combined with the primary antibody are immunostained with an antibody which is labeled by red fluorescence and can combine with the anti-CDX2 monoclonal antibody as a secondary antibody through the immunostaining process; and 3, whole nuclei of the cattle blastulae are subjected to fluorescent staining after completing the immunostaining to form the differential staining. On the basis of a case that CDX2 proteins specifically express in the trophoblastic cells and do not express in the inner cell mass cells, the method of the invention allows the differential staining to be carried out based on the red immunostaining of the CDX2 molecules and the blue staining of DAPI nuclei, so the accuracy is 100%, and obtained pictures are intelligible and beautiful.

Description

technical field [0001] The invention belongs to the field of mammalian embryology and relates to a differential staining method, in particular to a differential staining method for bovine blastocyst inner cell mass cells and trophoblast cells. Background technique [0002] The number of cells in the inner cell mass and the number of trophoblast cells in the blastocyst and their ratio are one of the criteria to measure the quality of the embryo. The staining that reflects the two cells in the blastocyst is called differential staining. The principle of the traditional differential staining method is based on the inner cell mass cells inside and the trophoblast cells outside, and red PI is used for 20s to stain the peripheral trophoblast cells, and then all the cells are stained with blue Hoechst33342, two pictures After merging, the pink peripheral cells are trophoblast cells, and the blue inner cells are inner cell mass cells. [0003] This traditional method of differenti...

AI Technical Summary

Problems solved by technology

[0003] This traditional method of differential staining of blastocysts is based on the above-mentioned simple principle of inside and outside positions, so the accuracy of differential staining is low, and the traditional differential staining still has the problem of unclear pictures

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