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

基本信息

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
项目类别：
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

项目摘要

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.

抽象的最新数据显示，已执行约 150 万次辅助生殖技术 (ART) 周期全世界每年估计有 350,000 名婴儿出生，这是 IVF 过程中的关键步骤之一。过程是选择高质量的胚胎进行子宫移植，目前这种选择很大程度上是基于的。确定囊胚的形态学标准和物理特征，而这些标准有。被证明有助于提高着床率、评估个体的生殖潜力因此，IVF 中心经常同时进行多个胚胎移植。这可能导致多胎妊娠，从而增加早产和死亡或终生的风险随着全球辅助生殖周期数量的增加，新生儿的残疾情况也有所改善。我们迫切需要开发更加定性和客观的预测胚胎活力的能力。更安全、更快速地评估胚胎质量和活力的方法可以提供重要的帮助 IVF 技术的进步是通过单胎胚胎移植而不是多个胚胎植入鉴于形态学评估的局限性，以增加成功妊娠的可能性。已经探索了多种评估胚胎活力的技术。测量胚胎培养基中的代谢物以及基因组和蛋白质组分析胚胎本身的光谱方法也被用来测量胚胎的数量。然而，这些方法是在植入前发育过程中产生的代谢物。消耗并需要训练有素的人员来分析复杂的数据。应用相量 FLIM（荧光寿命成像显微镜）方法，这是一种“非侵入式实时成像方法能够测量活体中的内源性自发荧光代谢物该方法捕获正在进行体外培养的胚胎的代谢能量来源的信息。被植入前胚胎用作胚胎质量和活力的读数。