Determination of the number of cells in preimplantation embryos by using noninvasive optical quadrature microscopy in conjunction with differential interference contrast microscopy.
Determination of the number of cells in preimplantation embryos by using noninvasive optical quadrature microscopy in conjunction with differential interference contrast microscopy.
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使用无创光学正交显微镜结合微分干涉对比显微镜测定植入前胚胎中的细胞数量。
DOI：
10.1017/s1431927607070171
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发表时间：
2007
期刊：
影响因子：
--
通讯作者：
Warner,CarolM
中科院分区：
文献类型：
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作者：
Newmark,JudithA;Warger2nd,WilliamC;Chang,ChihChing;Herrera,GustavoE;Brooks,DanaH;DiMarzio,CharlesA;Warner,CarolM
植入前胚胎中的细胞数量与胚胎的健康和活力直接相关。目前还没有非侵入性的方法来计数晚期植入前胚胎中的细胞数量。我们评估的能力，光学正交显微镜（OQM）计数的细胞数在小鼠植入前胚胎非侵入性。首先，为了测试可能的光毒性，我们将两细胞小鼠胚胎暴露于OQM和微分干涉对比（DIC）显微镜下，并评估其发育到囊胚阶段的能力。我们发现，从任何模式的显微镜下，长达2小时的曝光没有抑制发展。我们还利用OQM对8-桑椹期小鼠植入前胚胎进行了成像，并建立了两种细胞计数方法。轮廓签名法（CSM）仅使用OQM图像，相位相减法（PSM）使用OQM和DIC图像。我们将这两种方法与标准细胞计数技术进行了比较，发现PSM上级所有其他非侵入性细胞计数方法。我们在小鼠胚胎上的工作应该适用于人类胚胎。正确计数人类植入前胚胎中细胞数量的能力可能会导致体外受精（IVF）诊所中移植更少的胚胎，从而降低高风险多胞胎出生率。
The number of cells in a preimplantation embryo is directly correlated to the health and viability of the embryo. There are currently no methods to count the number of cells in late-stage preimplantation embryos noninvasively. We assessed the ability of optical quadrature microscopy (OQM) to count the number of cells in mouse preimplantation embryos noninvasively. First, to test for possible light toxicity, we exposed two-cell mouse embryos to OQM and differential interference contrast (DIC) microscopy and assessed their ability to develop to the blastocyst stage. We found no inhibition of development from either mode of microscopy for up to 2 h of light exposure. We also imaged eight-cell to morula stage mouse preimplantation embryos by OQM nd developed two methods for counting the number of cells. The contour signature method (CSM) used OQM images alone and the phase subtraction method (PSM) used both OQM and DIC images. We compared both methods to standard cell counting techniques and found that the PSM was superior to all other noninvasive cell counting methods. Our work on mouse embryos should be applicable to human embryos. The ability to correctly count the number of cells in human preimplantation embryos could lead to the transfer of fewer embryos in in vitro fertilization (IVF) clinics and consequently a lower rate of high-risk multiple-infant births.