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Information Journal Paper

Title

NOVEL APPROACH OF DIFFERENTIAL STAINING TO DETECT NECROTIC CELLS IN PREIMPLANTATION EMBRYOS

Pages

  103-106

Abstract

 Background: This novel approach describes a rapid and simple method for identification of necrotic vs. viable cells within a MAMMALIAN BLASTOCYST.Materials and Methods: Hatched bovine blastocysts produced in vitro were first incubated for 30 min in pre-equilibrated culture medium containing PROPIDIUM IODIDE (PI; 300mg/ml) and bisbenzimide (Hoechst: H33342; 5 mg/ml) fluorescent dyes. Embryos were then freed from residual dyes by thoroughly washing in warm phosphate buffer saline free of calcium and magnesium (PBS-), fixed in 2.5% glutharaldehyde and washed again in PBS- Stained embryos afterwards were mounted in a drop of glycerol over a microscopic slide. Prepared samples were examined under an epifluorescent microscope using the same excitation wavelength (330-385nm) and barrier filter (400nm) to distinguish necrosed vs. viable blastomers as being appeared in red and blue, respectively.Results: Obtained results showed that in cells with altered cell membrane such as late apoptotic or necrotic cells, PI and H33342 readily enter through the cytoplasmic barriers and so the chromatin materials are stained by both, but since PI quenches bisbenzimide fluorescence, necrotic blastomeres are seen in red to pinky red, while live cells are seen just as blue.Conclusion: Obtained results clearly indicated that this novel approach can be used as a simple, feasible and precise method for every embryology lab and with all the MAMMALIAN BLASTOCYSTs produced either in vitro or in vivo. The basic assay can be completed in 60 min, and valuable and reliable information can be obtained about the quality of the embryos.

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    • Cite

    APA: Copy

    HOSSEINI, SEYED MORTEZA, HAJIAN, MAHDI, ASGARI, VAJIHEH, FOROUZANFAR, MOHSEN, ABEDI, PARVANEH, & NASRESFAHANI, MOHAMMAD HOSSEIN. (2007). NOVEL APPROACH OF DIFFERENTIAL STAINING TO DETECT NECROTIC CELLS IN PREIMPLANTATION EMBRYOS. INTERNATIONAL JOURNAL OF FERTILITY AND STERILITY, 1(3), 103-106. SID. https://sid.ir/paper/304415/en

    Vancouver: Copy

    HOSSEINI SEYED MORTEZA, HAJIAN MAHDI, ASGARI VAJIHEH, FOROUZANFAR MOHSEN, ABEDI PARVANEH, NASRESFAHANI MOHAMMAD HOSSEIN. NOVEL APPROACH OF DIFFERENTIAL STAINING TO DETECT NECROTIC CELLS IN PREIMPLANTATION EMBRYOS. INTERNATIONAL JOURNAL OF FERTILITY AND STERILITY[Internet]. 2007;1(3):103-106. Available from: https://sid.ir/paper/304415/en

    IEEE: Copy

    SEYED MORTEZA HOSSEINI, MAHDI HAJIAN, VAJIHEH ASGARI, MOHSEN FOROUZANFAR, PARVANEH ABEDI, and MOHAMMAD HOSSEIN NASRESFAHANI, “NOVEL APPROACH OF DIFFERENTIAL STAINING TO DETECT NECROTIC CELLS IN PREIMPLANTATION EMBRYOS,” INTERNATIONAL JOURNAL OF FERTILITY AND STERILITY, vol. 1, no. 3, pp. 103–106, 2007, [Online]. Available: https://sid.ir/paper/304415/en

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