Assessment of the phasor Fluorescence Lifetime Imaging Microscopy (FLIM) Approach in an animal model

相量荧光寿命成像显微镜 (FLIM) 方法在动物模型中的评估

• 批准号: 9396700
• 负责人: Ken W.Y. Cho
• 金额: $ 22.19万
• 依托单位: UNIVERSITY OF CALIFORNIA-IRVINE
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-08-01 至 2019-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9396700
• 关键词: Adopted; Affect; Algorithms; Animal Model; Animals; Assisted Reproductive Technology; Behavior; Biological Assay; Birth; Breeding; Cell Count; Cell Size; Cells; Cessation of life; Characteristics; Chemicals; Classification; Clinic; Complex; Culture Media; Culture Techniques; Data; Detection; Development; Development Plans; Embryo; Embryo Transfer; Embryonic Development; Embryonic Lethal Mutation; Energy-Generating Resources; Evaluation; Fertilization in Vitro; Future; Generations; Genetic; Genomics; Glucose; Guidelines; Human; Human Resources; In Vitro; Individual; Live Birth; Maintenance; Manuals; Measurement; Measures; Metabolic; Microscopic; Morphology; Multiple Pregnancy; Mus; Noise; Nutrient; Oocytes; Ovulation Induction; Oxygen; Performance; Phase; Physical shape; Physicians; Pregnancy; Premature Birth; Procedures; Process; Proteomics; Pyruvate; Reactive Oxygen Species; Reading; Retrieval; Risk; S Phase; Safety; Signal Transduction; Solid; Spectroscopy, Fourier Transform Infrared; System; Techniques; Technology; Testing; Time; Training; Woman; Work; assisted reproduction; base; blastocyst; disability; fluorescence lifetime imaging; imaging approach; implantation; improved; indexing; instrumentation; mutant; natural Blastocyst Implantation; neonate; novel strategies; oxidized lipid; preimplantation; reproductive; success

Abstract The latest figure is that around 1.5 million Assisted Reproductive Technology (ART) cycles are performed each year worldwide, with an estimated 350,000 babies born. One of the critical steps during the IVF process is the selection of high-quality embryos for uterine transfer. This selection is currently based largely on defined morphological criteria and physical characteristics of the blastocyst. While such criteria have proven to be useful in improving implantation rates, assessment of the reproductive potential of individual embryos is not sufficient. Therefore, IVF centers often perform simultaneous transfers of multiple embryos that can result in multiple pregnancies, thus increasing the risk of preterm delivery and the death or lifelong disability of neonates. As the number of assisted reproduction cycles worldwide is increasing, improvements in our ability to predict embryo viability is urgently needed. Development of more qualitative and objective means for assessing embryo quality and viability that are safer and faster could provide significant advances in IVF by enabling singleton embryo transfers rather than the implantation of multiple embryos in order to increase the likelihood of a successful pregnancy. Given the limitations of morphologic evaluation, several technologies have been explored for the assessment of embryo viability. These include the measurement of metabolites in embryonic culture media along with genomic and proteomic profiling of the embryos themselves. Spectroscopic approaches have also been utilized to measure the amount of metabolites that arise during pre-implantation development. However, these approaches are time- consuming and require highly-trained personnel to analyze the complex data. Here we describe the application of a phasor-FLIM (Fluorescence-Lifetime Imaging Microscopy) approach, which is a “non- invasive” live imaging approach capable of measuring endogenous autofluorescent metabolites within living embryos undergoing in vitro culturing. The approach captures information on the metabolic energy sources utilized by pre-implantation embryos as readout of embryo quality and viability.

抽象的 最新数字是，进行了约150万个辅助生殖技术（ART）周期 每年在全球范围内，估计有35万婴儿出生。 IVF期间的关键步骤之一 过程是用于子宫转移的高质量胚胎的选择。此选择目前主要基于 关于胚泡的定义形态标准和物理特征。虽然这种标准有 被证明可用于提高实施率，评估个人的复制潜力 胚胎还不够。因此，IVF中心经常进行多个胚胎的简单转移 这可能导致多次怀孕，从而增加了早产的风险和死亡或终生 新生儿的残疾。随着全球辅助繁殖周期的数量正在增加，改善了 在我们预测胚胎生存力的能力中，迫切需要。发展更定性和客观 评估安全和更快的胚胎质量和生存能力的方法 通过实现单胎胚胎转移而不是植入多个胚胎，在IVF中的进步 为了增加成功怀孕的可能性。考虑到形态评估的局限性， 已经探索了几种技术以评估胚胎生存能力。这些包括 测量胚胎培养基中代谢物以及基因组和蛋白质组学分析的测量 胚胎本身。光谱方法也已用于测量 在植入前发育过程中出现的代谢产物。但是，这些方法是时间 - 消费并需要训练有素的人员来分析复杂数据。在这里，我们描述了 使用相flim（荧光千装的成像显微镜）方法的应用，这是“非 - 侵入性”实时成像方法，能够测量生活中内源性自动荧光代谢物 经过体外培养的胚胎。该方法捕获有关代谢能源的信息 通过植入前胚胎作为胚胎质量和生存能力的读数。