拉曼光谱检测在辅助生殖领域的研究进展
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摘要
辅助生殖技术(ART)的体外受精/胞浆内单精子注射-胚胎移植(IVF/ICSI-ET)需要实验室条件下处理两种配子、体外受精、体外培养形成胚胎或囊胚并挑选质量好的胚胎进行移植,使母亲妊娠,是目前解决不孕不育最有效的手段。因此,客观评估配子及植入前胚胎的质量显得非常重要。拉曼光谱不受被测物形态限制,无需样本制备或标记等复杂操作,属于无损伤检测。由于其具有检测迅速、耗时短、敏感度高及分析准确等特点,因此近年被广泛应用于快速检测蛋白质、核酸和脂类等生物大分子损伤。拉曼光谱技术主要有表面增强拉曼光谱技术、共焦显微拉曼光谱技术和傅里叶变换拉曼光谱技术3种。本文概述拉曼光谱的特点及上述3种主要拉曼光谱技术在生物样品分析方面的应用,探讨拉曼光谱技术应用于对精液、卵子和植入前胚胎质量检测的可能性,在辅助生殖领域具有潜在的应用前景。
Assisted reproductive technology(ART) is the most effective therapy for those infertile couples.In ART therapy such as IVF/ICSI-ET, both sperm and egg should be treated and fertilized in vitro. The fertilized ovum was then cultured in vitro, and the high-quality embryo was singled out for transfer. Therefore, it is very important how to evaluate the quality of sperm, egg and pre-transfer embryos. Raman spectroscopy is a non-damage detection method with the characteristics of speedliness, high sensitivity and accuracy, because the target object is not previously treated and marked. Raman spectroscopy has been used in the rapid detection of damage of biological macromolecule such as protein, nucleic acid and lipid in recent years. There are three kinds of Raman spectroscopy technologies: surface-enhanced Raman spectroscopy, confocal Raman spectroscopy and Fourier transform Raman spectroscopy. This article briefly discussed the characteristics of Raman spectroscopy and the possible application in ART for the evaluation of quality of semen, ovum and embryo, which provided us a reference for the clinical application in future.
Assisted reproductive technology(ART) is the most effective therapy for those infertile couples.In ART therapy such as IVF/ICSI-ET, both sperm and egg should be treated and fertilized in vitro. The fertilized ovum was then cultured in vitro, and the high-quality embryo was singled out for transfer. Therefore, it is very important how to evaluate the quality of sperm, egg and pre-transfer embryos. Raman spectroscopy is a non-damage detection method with the characteristics of speedliness, high sensitivity and accuracy, because the target object is not previously treated and marked. Raman spectroscopy has been used in the rapid detection of damage of biological macromolecule such as protein, nucleic acid and lipid in recent years. There are three kinds of Raman spectroscopy technologies: surface-enhanced Raman spectroscopy, confocal Raman spectroscopy and Fourier transform Raman spectroscopy. This article briefly discussed the characteristics of Raman spectroscopy and the possible application in ART for the evaluation of quality of semen, ovum and embryo, which provided us a reference for the clinical application in future.
引文
[1]Nishimune Y,T anaka H.Infertility caused by polymorphisms or mutations in spermatogenesis-specific genes[J].J Androl,2006,27(3):326-334.
[2]Sauer MV.Reproduction at an advanced maternal age and maternal health[J].Fertil Steril,2015,103(5):1136-1143.
[3]Boudjenah R,Molina-Gomes D,Torre A,et al.Associations between Individual and Combined Polymorphisms of the TNF and VEGF Genes and the Embryo Implantation Rate in Patients Undergoing In Vitro Fertilization(IVF)Programs[J].PLo S One,2014,9(9):e108287.
[4]de Mouzon J,Goossens V,Bhattacharya S,et al.Assisted reproductive technology in Europe,2007:results generated from European registers by ESHRE[J].Hum Reprod,2012,27(4):954-966.
[5]阿迪力·伊敏,艾尼瓦尔·吾术尔,沙依甫加马力·达吾来提.拉曼光谱的研究进展[J].西部皮革,2017,39(12):6-7.
[6]De Luca AC,Dholakia K,Mazilu M.Modulated Raman Spectroscopy for Enhanced Cancer Diagnosis at the Cellular Level[J].Sensors(Basel),2015,15(6):13680-13704.
[7]Kim HH.Endoscopic Raman Spectroscopy for Molecular Fingerprinting of Gastric Cancer:Principle to Implementation[J].Biomed Res Int,2015,2015:670121.
[8]Fleischmann M,Hendra PJ,Mc Quillan AJ.Raman spectra of pyridine adsorbed at a silver electrode[J].Chem Phys Lett,1974,26(2):163-166.
[9]张会敏,曲新艳,周喆,等.基于表面增强拉曼光谱的小鼠流感血清检测技术[J].实用医学杂志,2018,34(5):712-716.
[10]Witkowska E,Korsak D,Kowalska A,et al.Surface-enhanced Raman spectroscopy introduced into the International Standard Organization(ISO)regulations as an alternative method for detection and identification of pathogens in the food industry[J].Anal Bioanal Chem,2017,409(6):1555-1567.
[11]Kang JW,So PT,Dasari RR,et al.High resolution live cell Raman imaging using subcellular organelle-targeting SERS-sensitive gold nanoparticles with highly narrow intra-nanogap[J].Nano Lett,2015,15(3):1766-1772.
[12]Lee CW,Tseng FG.Surface enhanced Raman scattering(SERS)based biomicrofluidics systems for trace protein analysis[J].Biomicrofluidics,2018,12(1):011502.
[13]Bhamidipati M,Fabris L.Multiparametric Assessment of Gold Nanoparticle Cytotoxicity in Cancerous and Healthy Cells:The Role of Size,Shape,and Surface Chemistry[J].Bioconjug Chem,2017,28(2):449-460.
[14]Puppels GJ,de Mul FF,Otto C,et al.Studying single living cells and chromosomes by confocal Raman microspectroscopy[J].Nature,1990,347(6290):301-303.
[15]李书成,王允,崔晗,等.具有抗漂移特性的激光共焦拉曼光谱成像技术与系统[J].光谱学与光谱分析,2017,37(10):3249-3254.
[16]钟会清,张武,侯雨晴,等.激光共焦显微拉曼光谱技术在人舌鳞癌细胞检测中的应用[J].中国医学物理学杂志,2017,34(7):753-756.
[17]黄蓉,杨永健.拉曼光谱在药物分析中的研究进展[J].医药导报,2018,37(1):81-84.
[18]陈和生,孙育斌.几种塑料的傅里叶变换拉曼光谱分析[J].塑料科技,2012,40(6):69-72.
[19]Mansano ESB,de Morais GR,Moratto EM,et al.Combination of Histopathology and FT-Raman Spectroscopy for the Study of Experimental Paracoccidioidomycosis in the Spleen[J].Photochem Photobiol,2018,94(1):88-94.
[20]Bogliolo L,Murrone O,Piccinini M,et al.Evaluation of the impact of vitrification on the actin cytoskeleton of in vitro matured ovine oocytes by means of Raman microspectroscopy[J].J Assist Reprod Genet,2015,32(2):185-193.
[21]Rusciano G,De Canditiis C,Zito G,et al.Raman-microscopy investigation of vitrification-induced structural damages in mature bovine oocytes[J].PLo S One,2017,12(5):e0177677.
[22]Davidson B,Murray AA,Elfick A,et al.Raman microspectroscopy can be used to investigate the developmental stage of the mouse oocyte[J].PLo S One,2013,8(7):e67972.
[23]Bradley J,Pope I,Masia F,et al.Quantitative imaging of lipids in live mouse oocytes and early embryos using CARS microscopy[J].Development,2016,143(12):2238-2247.
[24]Bogliolo L,Murrone O,Di Emidio G,et al.Raman spectroscopybased approach to detect aging-related oxidative damage in the mouse oocyte[J].J Assist Reprod Genet,2013,30(7):877-882.
[25]Nagy ZP,Sakkas D,Behr B.Symposium:innovative techniques in human embryo viability assessment.Non-invasive assessment of embryo viability by metabolomic profiling of culture media(′metabolomics′)[J].Reprod Biomed Online,2008,17(4):502-507.
[26]Sikirzhytski V,Sikirzhytskaya A,Lednev IK.Multidimensional Raman spectroscopic signatures as a tool for forensic identification of body fluid traces:a review[J].Appl Spectrosc,2011,65(11):1223-1232.
[27]李征,陈美佳,钟惠芝,等.不同精液参数对体外受精结局的影响及预测价值[J].广西医学,2017,39(6):885-887.
[28]Gilany K,Moazeni-Pourasil RS,Jafarzadeh N,et al.Metabolomics fingerprinting of the human seminal plasma of asthenozoospermic patients[J].Mol Reprod Dev,2014,81(1):84-86.
[29]Huang Z,Chen X,Li Y,et al.Quantitative determination of citric acid in seminal plasma by using Raman spectroscopy[J].Appl Spectrosc,2013,67(7):757-760.
[30]Chen X,Huang Z,Feng S,et al.Analysis and differentiation of seminal plasma via polarized SERS spectroscopy[J].Int J Nanomedicine,2012,7:6115-6121.
[31]刘锋,朱勇,李铮.基于拉曼光谱评估人类精子存活能力的研究[J].生殖与避孕,2016,36(2):156-159.
[32]Sánchez V,Redmann K,Wistuba J,et al.Oxidative DNA damage in human sperm can be detected by Raman microspectroscopy[J].Fertil Steril,2012,98(5):1124-1129.e1-e3.
[33]Liu F,Zhu Y,Liu Y,et al.Real-time Raman microspectroscopy scanning of the single live sperm bound to human zona pellucida[J].Fertil Steril,2013,99(3):684-689.e4.
[34]Majumdar G,Majumdar A,Verma IC,et al.Relationship Between Morphology,Euploidy and Implantation Potential of Cleavage and Blastocyst Stage Embryos[J].J Hum Reprod Sci,2017,10(1):49-57.
[35]Bronet F,Agudo D.Should we forget about embryos till day 5?[J].Curr Opin Obstet Gynecol,2017,29(3):107-111.
[36]Fabian D,Ka cˇmarováM,KubandováJ,et al.Raman spectroscopy analysis of differences in composition of spent culture media of in vitro cultured preimplantation embryos isolated from normal and fat mice dams[J].Reprod Biol,2016,16(2):120-129.
[37]Li Q,Suasnavas E,Xiao L,et al.Label-free and non-invasive monitoring of porcine trophoblast derived cells:differentiation in serum and serum-free media[J].J Biophotonics,2015,8(8):638-645.
[38]Gardner DK,Lane M,Stevens J,et al.Noninvasive assessment of human embryo nutrient consumption as a measure of developmental potential[J].Fertil Steril,2001,76(6):1175-1180.
[39]Gardner DK,Leese HJ.Non-invasive measurement of nutrient uptake by single cultured pre-implantation mouse embryos[J].Hum Reprod,1986,1(1):25-27.
[40]Conaghan J,Hardy K,Handyside AH,et al.Selection criteria for human embryo transfer:a comparison of pyruvate uptake and morphology[J].J Assist Reprod Genet,1993,10(1):21-30.
[41]Houghton FD,Hawkhead JA,Humpherson PG,et al.Non-invasive amino acid turnover predicts human embryo developmental capacity[J].Hum Reprod,2002,17(4):999-1005.
[42]Brison DR,Houghton FD,Falconer D,et al.Identification of viable embryos in IVF by non-invasive measurement of amino acid turnover[J].Hum Reprod,2004,19(10):2319-2324.
[43]Huang Z,Sun Y,Wang J,et al.Rapid and nondestructive method for evaluation of embryo culture media using drop coating deposition Raman spectroscopy[J].J Biomed Opt,2013,18(12):127003.
[44]Ding J,Xu T,Tan X,et al.Raman spectrum:A potential biomarker for embryo assessment during in vitro fertilization[J].Exp Ther Med,2017,13(5):1789-1792.
[45]Ishigaki M,Hashimoto K,Sato H,et al.Non-destructive monitoring of mouse embryo development and its qualitative evaluation at the molecular level using Raman spectroscopy[J].Sci Rep,2017,7:43942.
[46]Seli E,Sakkas D,Scott R,et al.Noninvasive metabolomic profiling of embryo culture media using Raman and near-infrared spectroscopy correlates with reproductive potential of embryos in women undergoing in vitro fertilization[J].Fertil Steril,2007,88(5):1350-1357.
[47]Scott R,Seli E,Miller K,et al.Noninvasive metabolomic profiling of human embryo culture media using Raman spectroscopy predicts embryonic reproductive potential:a prospective blinded pilot study[J].Fertil Steril,2008,90(1):77-83.
[48]Zhao Q,Yin T,Peng J,et al.Noninvasive metabolomic profiling of human embryo culture media using a simple spectroscopy adjunct to morphology for embryo assessment in in vitro fertilization(IVF)[J].Int J Mol Sci,2013,14(4):6556-6570.
[49]Shen AG,Peng J,Zhao QH,et al.Accurate and noninvasive embryos screening during in vitro fertilization(IVF)assisted by Raman analysis ofembryos culture medium[J].Laser Phys Lett,2012,9(4):322-328.
[50]Bastu E,Parlatan U,Basar G,et al.Spectroscopic analysis of embryo culture media for predicting reproductive potential in patients undergoing in vitro fertilization[J].Turk J Obstet Gynecol,2017,14(3):145-150.
[2]Sauer MV.Reproduction at an advanced maternal age and maternal health[J].Fertil Steril,2015,103(5):1136-1143.
[3]Boudjenah R,Molina-Gomes D,Torre A,et al.Associations between Individual and Combined Polymorphisms of the TNF and VEGF Genes and the Embryo Implantation Rate in Patients Undergoing In Vitro Fertilization(IVF)Programs[J].PLo S One,2014,9(9):e108287.
[4]de Mouzon J,Goossens V,Bhattacharya S,et al.Assisted reproductive technology in Europe,2007:results generated from European registers by ESHRE[J].Hum Reprod,2012,27(4):954-966.
[5]阿迪力·伊敏,艾尼瓦尔·吾术尔,沙依甫加马力·达吾来提.拉曼光谱的研究进展[J].西部皮革,2017,39(12):6-7.
[6]De Luca AC,Dholakia K,Mazilu M.Modulated Raman Spectroscopy for Enhanced Cancer Diagnosis at the Cellular Level[J].Sensors(Basel),2015,15(6):13680-13704.
[7]Kim HH.Endoscopic Raman Spectroscopy for Molecular Fingerprinting of Gastric Cancer:Principle to Implementation[J].Biomed Res Int,2015,2015:670121.
[8]Fleischmann M,Hendra PJ,Mc Quillan AJ.Raman spectra of pyridine adsorbed at a silver electrode[J].Chem Phys Lett,1974,26(2):163-166.
[9]张会敏,曲新艳,周喆,等.基于表面增强拉曼光谱的小鼠流感血清检测技术[J].实用医学杂志,2018,34(5):712-716.
[10]Witkowska E,Korsak D,Kowalska A,et al.Surface-enhanced Raman spectroscopy introduced into the International Standard Organization(ISO)regulations as an alternative method for detection and identification of pathogens in the food industry[J].Anal Bioanal Chem,2017,409(6):1555-1567.
[11]Kang JW,So PT,Dasari RR,et al.High resolution live cell Raman imaging using subcellular organelle-targeting SERS-sensitive gold nanoparticles with highly narrow intra-nanogap[J].Nano Lett,2015,15(3):1766-1772.
[12]Lee CW,Tseng FG.Surface enhanced Raman scattering(SERS)based biomicrofluidics systems for trace protein analysis[J].Biomicrofluidics,2018,12(1):011502.
[13]Bhamidipati M,Fabris L.Multiparametric Assessment of Gold Nanoparticle Cytotoxicity in Cancerous and Healthy Cells:The Role of Size,Shape,and Surface Chemistry[J].Bioconjug Chem,2017,28(2):449-460.
[14]Puppels GJ,de Mul FF,Otto C,et al.Studying single living cells and chromosomes by confocal Raman microspectroscopy[J].Nature,1990,347(6290):301-303.
[15]李书成,王允,崔晗,等.具有抗漂移特性的激光共焦拉曼光谱成像技术与系统[J].光谱学与光谱分析,2017,37(10):3249-3254.
[16]钟会清,张武,侯雨晴,等.激光共焦显微拉曼光谱技术在人舌鳞癌细胞检测中的应用[J].中国医学物理学杂志,2017,34(7):753-756.
[17]黄蓉,杨永健.拉曼光谱在药物分析中的研究进展[J].医药导报,2018,37(1):81-84.
[18]陈和生,孙育斌.几种塑料的傅里叶变换拉曼光谱分析[J].塑料科技,2012,40(6):69-72.
[19]Mansano ESB,de Morais GR,Moratto EM,et al.Combination of Histopathology and FT-Raman Spectroscopy for the Study of Experimental Paracoccidioidomycosis in the Spleen[J].Photochem Photobiol,2018,94(1):88-94.
[20]Bogliolo L,Murrone O,Piccinini M,et al.Evaluation of the impact of vitrification on the actin cytoskeleton of in vitro matured ovine oocytes by means of Raman microspectroscopy[J].J Assist Reprod Genet,2015,32(2):185-193.
[21]Rusciano G,De Canditiis C,Zito G,et al.Raman-microscopy investigation of vitrification-induced structural damages in mature bovine oocytes[J].PLo S One,2017,12(5):e0177677.
[22]Davidson B,Murray AA,Elfick A,et al.Raman microspectroscopy can be used to investigate the developmental stage of the mouse oocyte[J].PLo S One,2013,8(7):e67972.
[23]Bradley J,Pope I,Masia F,et al.Quantitative imaging of lipids in live mouse oocytes and early embryos using CARS microscopy[J].Development,2016,143(12):2238-2247.
[24]Bogliolo L,Murrone O,Di Emidio G,et al.Raman spectroscopybased approach to detect aging-related oxidative damage in the mouse oocyte[J].J Assist Reprod Genet,2013,30(7):877-882.
[25]Nagy ZP,Sakkas D,Behr B.Symposium:innovative techniques in human embryo viability assessment.Non-invasive assessment of embryo viability by metabolomic profiling of culture media(′metabolomics′)[J].Reprod Biomed Online,2008,17(4):502-507.
[26]Sikirzhytski V,Sikirzhytskaya A,Lednev IK.Multidimensional Raman spectroscopic signatures as a tool for forensic identification of body fluid traces:a review[J].Appl Spectrosc,2011,65(11):1223-1232.
[27]李征,陈美佳,钟惠芝,等.不同精液参数对体外受精结局的影响及预测价值[J].广西医学,2017,39(6):885-887.
[28]Gilany K,Moazeni-Pourasil RS,Jafarzadeh N,et al.Metabolomics fingerprinting of the human seminal plasma of asthenozoospermic patients[J].Mol Reprod Dev,2014,81(1):84-86.
[29]Huang Z,Chen X,Li Y,et al.Quantitative determination of citric acid in seminal plasma by using Raman spectroscopy[J].Appl Spectrosc,2013,67(7):757-760.
[30]Chen X,Huang Z,Feng S,et al.Analysis and differentiation of seminal plasma via polarized SERS spectroscopy[J].Int J Nanomedicine,2012,7:6115-6121.
[31]刘锋,朱勇,李铮.基于拉曼光谱评估人类精子存活能力的研究[J].生殖与避孕,2016,36(2):156-159.
[32]Sánchez V,Redmann K,Wistuba J,et al.Oxidative DNA damage in human sperm can be detected by Raman microspectroscopy[J].Fertil Steril,2012,98(5):1124-1129.e1-e3.
[33]Liu F,Zhu Y,Liu Y,et al.Real-time Raman microspectroscopy scanning of the single live sperm bound to human zona pellucida[J].Fertil Steril,2013,99(3):684-689.e4.
[34]Majumdar G,Majumdar A,Verma IC,et al.Relationship Between Morphology,Euploidy and Implantation Potential of Cleavage and Blastocyst Stage Embryos[J].J Hum Reprod Sci,2017,10(1):49-57.
[35]Bronet F,Agudo D.Should we forget about embryos till day 5?[J].Curr Opin Obstet Gynecol,2017,29(3):107-111.
[36]Fabian D,Ka cˇmarováM,KubandováJ,et al.Raman spectroscopy analysis of differences in composition of spent culture media of in vitro cultured preimplantation embryos isolated from normal and fat mice dams[J].Reprod Biol,2016,16(2):120-129.
[37]Li Q,Suasnavas E,Xiao L,et al.Label-free and non-invasive monitoring of porcine trophoblast derived cells:differentiation in serum and serum-free media[J].J Biophotonics,2015,8(8):638-645.
[38]Gardner DK,Lane M,Stevens J,et al.Noninvasive assessment of human embryo nutrient consumption as a measure of developmental potential[J].Fertil Steril,2001,76(6):1175-1180.
[39]Gardner DK,Leese HJ.Non-invasive measurement of nutrient uptake by single cultured pre-implantation mouse embryos[J].Hum Reprod,1986,1(1):25-27.
[40]Conaghan J,Hardy K,Handyside AH,et al.Selection criteria for human embryo transfer:a comparison of pyruvate uptake and morphology[J].J Assist Reprod Genet,1993,10(1):21-30.
[41]Houghton FD,Hawkhead JA,Humpherson PG,et al.Non-invasive amino acid turnover predicts human embryo developmental capacity[J].Hum Reprod,2002,17(4):999-1005.
[42]Brison DR,Houghton FD,Falconer D,et al.Identification of viable embryos in IVF by non-invasive measurement of amino acid turnover[J].Hum Reprod,2004,19(10):2319-2324.
[43]Huang Z,Sun Y,Wang J,et al.Rapid and nondestructive method for evaluation of embryo culture media using drop coating deposition Raman spectroscopy[J].J Biomed Opt,2013,18(12):127003.
[44]Ding J,Xu T,Tan X,et al.Raman spectrum:A potential biomarker for embryo assessment during in vitro fertilization[J].Exp Ther Med,2017,13(5):1789-1792.
[45]Ishigaki M,Hashimoto K,Sato H,et al.Non-destructive monitoring of mouse embryo development and its qualitative evaluation at the molecular level using Raman spectroscopy[J].Sci Rep,2017,7:43942.
[46]Seli E,Sakkas D,Scott R,et al.Noninvasive metabolomic profiling of embryo culture media using Raman and near-infrared spectroscopy correlates with reproductive potential of embryos in women undergoing in vitro fertilization[J].Fertil Steril,2007,88(5):1350-1357.
[47]Scott R,Seli E,Miller K,et al.Noninvasive metabolomic profiling of human embryo culture media using Raman spectroscopy predicts embryonic reproductive potential:a prospective blinded pilot study[J].Fertil Steril,2008,90(1):77-83.
[48]Zhao Q,Yin T,Peng J,et al.Noninvasive metabolomic profiling of human embryo culture media using a simple spectroscopy adjunct to morphology for embryo assessment in in vitro fertilization(IVF)[J].Int J Mol Sci,2013,14(4):6556-6570.
[49]Shen AG,Peng J,Zhao QH,et al.Accurate and noninvasive embryos screening during in vitro fertilization(IVF)assisted by Raman analysis ofembryos culture medium[J].Laser Phys Lett,2012,9(4):322-328.
[50]Bastu E,Parlatan U,Basar G,et al.Spectroscopic analysis of embryo culture media for predicting reproductive potential in patients undergoing in vitro fertilization[J].Turk J Obstet Gynecol,2017,14(3):145-150.