本文關(guān)鍵詞：人植入前胚胎細(xì)胞超薄電鏡切片的制作及應(yīng)用TUNEL技術(shù)對該細(xì)胞凋亡的原位檢測　出處：《華中科技大學(xué)》2006年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 人植入前胚胎 超薄切片 透射電鏡 原位末端標(biāo)記法 人植入前胚胎 凋亡

【摘要】： 目的:改進(jìn)人植入前胚胎細(xì)胞超薄切片的制備方法,觀察不同時(shí)期人早期胚胎細(xì)胞超微結(jié)構(gòu)的變化。 方法:選取我院生殖中心試管嬰兒助孕技術(shù)后異常受精的3原核胚胎,于2-8細(xì)胞期進(jìn)行固定,改進(jìn)透射電鏡的常規(guī)操作方法,使用3%～4%瓊脂進(jìn)行預(yù)包埋,制成超薄切片后在鏡下觀察植入前胚胎細(xì)胞凋亡的超微結(jié)構(gòu)。 結(jié)果:在鏡下可見胚胎細(xì)胞正常形態(tài)和凋亡的形態(tài)變化。 結(jié)論:使用本方法可以進(jìn)行少量胚胎的超薄切片。 目的:建立原位末端標(biāo)記法(TdT-mediated nick end labeling TUNEL)檢測人植入前胚胎凋亡征象的方法。 方法:取試管嬰兒助孕技術(shù)后異常受精的3原核胚胎,于2-10細(xì)胞期進(jìn)行固定,采用TUNEL法檢測3原核胚胎中的凋亡征象。 結(jié)果:TUNEL陽性信號(hào)為綠色強(qiáng)熒光,位于胞核,呈環(huán)狀或顆粒狀。在發(fā)育正常和發(fā)育停滯胚胎中,未檢測到凋亡征象;在有碎片的胚胎中20%可檢測到TUNEL陽性信號(hào)。 結(jié)論:采用TUNEL法加熒光共焦顯微鏡觀察凋亡細(xì)胞的方法可以有效檢測人植入前胚胎中的凋亡征象,其中細(xì)胞碎片的出現(xiàn)可能與凋亡有關(guān)。
[Abstract]:Aim: to improve the preparation of ultrathin sections of human preimplantation embryonic cells and observe the ultrastructural changes of human early embryonic cells at different stages. Methods: the 3 prokaryotic embryos of IVF in our hospital were fixed at 2-8 cell stage, and the conventional operation of transmission electron microscope was improved. The ultrastructure of apoptosis of preimplantation embryo cells was observed under microscope after ultrathin sections were made by using 3% Agar and 4% Agar. Results: the morphological changes of normal and apoptotic embryonic cells were observed under microscope. Conclusion: this method can be used to carry out ultrathin sections of a small number of embryos. Aim: to establish a TdT-mediated nick end labeling Tunel method for the detection of apoptosis in human preimplantation embryos. Methods: the 3 prokaryotic embryos with abnormal fertilization after IVF were fixed in 2-10 cell phase and the apoptotic signs in 3 prokaryotic embryos were detected by TUNEL method. Results the positive signal of TUNEL was green strong fluorescence, which was located in the nucleus and was annular or granular. No apoptotic sign was detected in normal and stunted embryos. TUNEL positive signals were detected in 20% of the embryos with fragments. Conclusion: the method of TUNEL and fluorescence confocal microscopy can be used to detect apoptosis in human preimplantation embryos, and the appearance of cell fragments may be related to apoptosis.
【學(xué)位授予單位】：華中科技大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2006
【分類號(hào)】：R321

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