血红细胞和斑马鱼的生物物理特性及检测方法
摘要
在科学技术飞速发展的今天,环境保护问题越来越受到人们的重视,随着日益严峻的环境等诸多问题的出现,各种疾病也随之而生,在这种情况下各种新药的研制也越来越普遍。
目前对于如何标度生物体的活性尚没有一种比较科学的方法,为了试图解决这一问题,本论文主要阐述两个方面的内容,即通过生物非线性的大小来表征生物体的活性大小,以及通过一种图像处理的方法观察不同环境下的斑马鱼的心脏健康性,以此来研究环境的影响。
理论和实验研究表明,相对于传统的光学的方法来研究非线性,我们这种交变电场的方法能够很方便的获得较多的数据,配合我们设计的硬件和软件,可以节约实验成本和所需时间。
斑马鱼的研究方面我们同样也自己设计了一套装置,可以适时采集动态图像,并配合我们的软件(最主要的是核心算法)可以准确的获取类似心电图的数据,对新药研制方面有很重要的贡献,目前这方面的成果我们已经申请国家发明专利。
We are trying to find a way to measure the characteristics of the biological systems, and we find two ways to detect it as the following.
When a suspension consisting of electric particles having nonlinear characteristics is subjected to a sinusoidal alternating current (ac) electric field, the electric response will generally consist of ac fields at frequencies of higher-order harmonics. We experimentally report on harmonic generation by erythrocytes subjected to an ac electric field. We find that both even and odd harmonics are sensitive to cell shapes, conductivities, field frequencies, and field magnitude. Theoretical analysis based on a phenomenological model yield predictions that are in excellent agreement with the experiments. Thus, it becomes possible to detect nonlinear characteristics, shapes, and conductivities of erythrocytes by measuring such ac responses.
On the other hand to develop an accurate and convenient method for measuring the heart rate of zebra fish in vivo, a system combining fast differential interference contrast (DIC) imaging with an autocorrelation technique is established. The imaging correlation coefficient corr(i, j) between frame i, selected from the obtained time-lapse imaging series as the reference image, and any other frame j, is calculated as the time-dependent cycle course. Heat rate is determined by the cycle period of the corr with a high temporal resolution of 4 ms, achieved by fast charge-coupled device (CCD) imaging of 250 frames per second. With this high-resolution system, we find that 1mg/L cadmium not only induces the slowing of the heart rate, but also caused signs of arrhythmia in treated fish.
目前对于如何标度生物体的活性尚没有一种比较科学的方法,为了试图解决这一问题,本论文主要阐述两个方面的内容,即通过生物非线性的大小来表征生物体的活性大小,以及通过一种图像处理的方法观察不同环境下的斑马鱼的心脏健康性,以此来研究环境的影响。
理论和实验研究表明,相对于传统的光学的方法来研究非线性,我们这种交变电场的方法能够很方便的获得较多的数据,配合我们设计的硬件和软件,可以节约实验成本和所需时间。
斑马鱼的研究方面我们同样也自己设计了一套装置,可以适时采集动态图像,并配合我们的软件(最主要的是核心算法)可以准确的获取类似心电图的数据,对新药研制方面有很重要的贡献,目前这方面的成果我们已经申请国家发明专利。
We are trying to find a way to measure the characteristics of the biological systems, and we find two ways to detect it as the following.
When a suspension consisting of electric particles having nonlinear characteristics is subjected to a sinusoidal alternating current (ac) electric field, the electric response will generally consist of ac fields at frequencies of higher-order harmonics. We experimentally report on harmonic generation by erythrocytes subjected to an ac electric field. We find that both even and odd harmonics are sensitive to cell shapes, conductivities, field frequencies, and field magnitude. Theoretical analysis based on a phenomenological model yield predictions that are in excellent agreement with the experiments. Thus, it becomes possible to detect nonlinear characteristics, shapes, and conductivities of erythrocytes by measuring such ac responses.
On the other hand to develop an accurate and convenient method for measuring the heart rate of zebra fish in vivo, a system combining fast differential interference contrast (DIC) imaging with an autocorrelation technique is established. The imaging correlation coefficient corr(i, j) between frame i, selected from the obtained time-lapse imaging series as the reference image, and any other frame j, is calculated as the time-dependent cycle course. Heat rate is determined by the cycle period of the corr with a high temporal resolution of 4 ms, achieved by fast charge-coupled device (CCD) imaging of 250 frames per second. With this high-resolution system, we find that 1mg/L cadmium not only induces the slowing of the heart rate, but also caused signs of arrhythmia in treated fish.
引文
[1] B. L. McClain, I. J. Finkelstein, and M. D. Fayer, "Dynamics of Hemoglobin in Human Erythrocytes and in Solution: Influence of Viscosity Studied by Ultrafast Vibrational Echo Experiments" , J. Am. Chem. Soc. 126, 15702 (2004).
[2] L. Haider, P. Snabre, and M. Boynard, Biophys. "Rheology and Ultrasound Scattering from Aggregated Red Cell Suspensions in Shear Flow" , J. 87, 2322 (2004).
[3] D. R. Daniels, J. C. Wang, R. W. Briehl, and M. S. Turner, "Deforming biological membranes: How the cytoskeleton affects a polymerizing fiber" , J. Chem. Phys. 124, 024903 (2006).
[4] G. M. Grotenbreg et al., " β-Turn Modified Gramicidin S Analogues Containing Arylated Sugar Amino Acids Display Antimicrobial and Hemolytic Activity Comparable to the Natural Product" , J. Am. Chem. Soc. 128, 7559 (2006).
[5] E. J. Ding and C. K. Aidun, "Cluster Size Distribution and Scaling for Spherical Particles and Red Blood Cells in Pressure-Driven Flows at Small Reynolds Number" , Phys. Rev. Lett. 96, 204502 (2006).
[6] S. Kaufmann and M. Tanaka, "Cell Adhesion onto Highly Curved Surfaces: One-Step Immobilization of Human Erythrocyte Membranes on Silica Beads" , ChemPhysChem 4, 699 (2003).
[7] S. B. Rochal and V. L. Lorman, "Cytoskeleton Influence on Normal and Tangent Fluctuation Modes in the Red Blood Cells" , Phys. Rev. Lett. 96, 248102 (2006).
[8] L. C. L. Lin, N. Gov, and F. L. H. Brown, "Nonequilibrium membrane fluctuations driven by active proteins" , J. Chem. Phys. 124, 074903 (2006).
[9] R. D. Schaller, J. C. Johnson, and R. J. Saykally, "Time-Resolved Second Harmonic Generation Near-Field Scanning Optical Microscopy" , ChemPhysChem 4, 1243 (2003).
[10] J. P. Huang and K. W. Yu, J. "Nonlinear ac responses of electro-magnetorheological fluids" , Chem. Phys. 121, 7526 (2004).
[11] L. Gao, L. P. Gu, and Z. Y. Li, "Optical bistability and tristability in nonlinear metal/dielectric composite media of nonspherical particles" , Phys. Rev. E 68, 066601 (2003).
[12] T. B. Jones, Electromechanics of Particles (Cambridge University Press, New York, 1995).
[13] A. M. Woodward, A. Jones, and X. Zhang, "Rapid and non-invasive quantification of metabolic substrates in biological cell suspensions using non—linear dielectric spectroscopy with multivariate calibration and artificial neural networks. Principles and applications " , Bioelectrochem. Bioenerg. 40, 99 (1996).
[14] N. Gov, "Membrane Undulations Driven by Force Fluctuations of Active Proteins" , Phys. Rev. Lett. 93, 268104 (2004).
[15] R. D. Astumian, "Adiabatic Pumping Mechanism for Ion Motive ATPases" , Phys. Rev. Lett. 91, 118102 (2003).
[16] D. Nawarathna, J. H. Miller, Jr., J. R. Claycomb, G. Cardenas, and D. Warmflash, "Harmonic Response of Cellular Membrane Pumps to Low Frequency Electric Fields" , Phys. Rev. Lett. 95, 158103 (2005).
[17] T. Y. Tsong and R. D. Astumian, "863 -Absorption and conversion of electric field energy by membrane bound atpases" , Bioelectrochem. Bioenerg. 15, 457 (1986).
[18] R. D. Astumian and I. Derenyi, "Fluctuation driven transport and models of molecular motors and pumps" , Eur. Biophys. J. 27, 474 (1998).
[19] B. Chanda, O. K. Asamoah, R. Blunck, B. Roux, and F. Bezanilla, "Gating charge displacement in voltage-gated ion channels involves limited transmembrane movement" , Nature (London) 436, 852 (2005).
[20] C. L. Davey, H. M. Davey, and D. B. Kell, "On the dielectric properties of cell suspensions at high volume fractions" , Bioelectrochem. Bioenerg. 28, 319 (1992).
[21] A. M. Woodward and D. B. Kell, "On the nonlinear dielectric properties of biological systems Saccharomyces cerevisiae" , Bioelectrochem. Bioenerg. 24, 83 (1990).
[22] Garnett, J. C.M. Colors in metal glasses and in metal films [J]. Philos. Trans. R. Soc. London Ser. A., 1904, 203 (2): 385-420.
[23] Garnett, J. C. M. Absorptance to Selected Averaging Procedures and Solar Irradiance Distributions [J]. Trans. R. Soc. London Ser. A, 1906, 205 (5) : 237-284.
[24] Bruggeman, D. A. G. Berechnung verschiedener physikalischer Konstanten von heterogenen Substanzen [J]. Ann. Phys. (Leipzig), 1935, 24 (8) :636-679.
[25] L. D. Landau, E. M. Lifshitz, and L. P. Pitaevskii, Electrodynamics of Continuous Media, 2nd ed. (Pergamon, New York, 1984), Chap. II.
[26] J. Gimsa and D. Wachner, " A polarization model overcoming the geometric restrictions of the Laplace solution for spheroidal cells: obtaining new equations for field-induced forces and transmembrane potential" , Biophys. J. 77, 1316 (1999).
[27] M. Pavlin and D. Miklavcic, "Effective conductivity of a suspension of permeabilized cells: a theoretical analysis" , Biophys. J. 85, 719 (2003).
[28] J. P. Huang and K. W. Yu, "Enhanced nonlinear optical responses of materials: Composite effects" , Phys. Rep. 431, 87 (2006).
[29] D. A. G. Bruggeman, Ann. Phys. 「Leipzig(?) 24, 636 (1935).
[30] D. StroudandP. M. Hui, "Nonlinear susceptibilities of granular matter" , Phys. Rev. B 37, 8719 (1988).
[31] M. C. Fishman, "Genomics—zebrafish—the canonical vertebrate" , Science 294(5545), 1290-1291 (2001).
[32] G. Lomberk, "Animal models" , Pancreatol. 6(5), 427-428 (2006).
[33] J. R. Hove, R. W. Koster, A. S. Forouhar, G. Acevedo-Bolton, S. E. Fraser, and M. Gharib, "Intracardiac fluid forces are an essential epigenetic factor for embryonic cardiogenesis" , Nature (London) 421(6919), 172-178 (2003).
[34] A. S. Forouhar and M. Liebling, "The embryonic vertebrate heart tube is a dynamic suction pump" , Science 312(5774), 751-753 (2006).
[35] DJ; Jones, IL; Ellinor, PT, et al., "In vivo recording of adult zebrafish electrocardiogram and assessment of drug-induced QT prolongation Milan" AMERICAN JOURNAL OF PHYSIOLOGY-HEART AND CIRCULATORY PHYSIOLOGY. 291(1), H269-H273 (2006).
[36] Burns, CG; Milan, DJ; Grande, EJ, et al. "High-throughput assay for small molecules that modulate zebrafish embryonic heart rate" , NATURE CHEMICAL BIOLOGY 1(5) 263-264 (2005).
[37] W. R. Barrionuevo and W. W. Burggren, "02 consumption and heart rate in developing zebrafish (Danio rerio): influence of temperature and ambient O2" , Am. J. Physiol. Regulatory Integrative Comp. Physiol. 276, 505-513(1999).
[38] E: Jacob, M. Drexel, T. Schwerte, and B. Pelster, "Influence of hypoxia and of hypoxemia on the development of cardiac activity in zebrafish larvae" , Am. J. Physiol. Regulatory Integrative Comp. Physiol. 283, 911-917 (2002).
[39] C. G. Burns, D. J. Milan, E. J. Grande, W. Rottbauer, C. A. MacRae, and M. C. Fishman, "High-throughput assay for small molecules that modulate zebrafish embryonic heart rate" , Nat. Chem. Biol. 1(5), 263-264 (2005).
[40] M. Westerfield, The Zebrafish Book, Chap. 2, Eugene Press, Eugene, OR (1995).
[41] V. Viasnoff, F. Lequeux, and D. J. Pine, "Multispeckle diffusingwave spectroscopy: a tool to study slow relaxation and timedependent dynamics" , Rev. Sci. Instrum. 73(6), 2336-2344 (2002).
[42] A. V. Hallare, M. Schirling, T. Luckenbach, H. R. Kohler, and R. Triebskorn, "Combined effects of temperature and cadmium on developmental parameters and biomarker responses in zebrafish (Danio rerio) embryos", J. Therm. Biol. 30(1), 7-17 (2005). [43] Muller, M. ; Squier, J. A. ; Wilson, K. R. ; Brakenhoff, G. J. J. Microsc. 1998,191,266-74.
[44]Squier,J.A.;Muller,M.Appl.Opt.1999,38,5789-94.
[45]Richard D.Schaller,Justin C.Johnson,and Richard J.Saykally,"Nonlinear Chemical Imaging Microscopy;Near-Field Third Harmonic Generation Imaging of Human Red Blood Cells",Analytical Chemistry,72(21),5361-5364(2000).
[46]TSANG,T.Y.F,"OPTICAL 3RD-HARMONIC GENERATION AT INTERFACES",PHYSICAL REVIEW A,52(5) 4116-4125(1995).
[47]Barad,Y.;Eisenberg,H.;Horowitz,M.;Silberberg,Y.Appl.Phys.Lett.1997,70,922-4.
[48]Squier,J.A.;Muller,M.;Brakenhoff,G.J.;Wilson,K.R.Opt.Express 1998,3,315-24.
[49]Yelin,D.;Silberberg,Y.Opt.Express 1999,5,169-75.
[50]Yelin,D.;Silberberg,Y.;Barad,Y.;Patel,J.S.Appl.Phys.Lett:1999,74,3107-9.
[2] L. Haider, P. Snabre, and M. Boynard, Biophys. "Rheology and Ultrasound Scattering from Aggregated Red Cell Suspensions in Shear Flow" , J. 87, 2322 (2004).
[3] D. R. Daniels, J. C. Wang, R. W. Briehl, and M. S. Turner, "Deforming biological membranes: How the cytoskeleton affects a polymerizing fiber" , J. Chem. Phys. 124, 024903 (2006).
[4] G. M. Grotenbreg et al., " β-Turn Modified Gramicidin S Analogues Containing Arylated Sugar Amino Acids Display Antimicrobial and Hemolytic Activity Comparable to the Natural Product" , J. Am. Chem. Soc. 128, 7559 (2006).
[5] E. J. Ding and C. K. Aidun, "Cluster Size Distribution and Scaling for Spherical Particles and Red Blood Cells in Pressure-Driven Flows at Small Reynolds Number" , Phys. Rev. Lett. 96, 204502 (2006).
[6] S. Kaufmann and M. Tanaka, "Cell Adhesion onto Highly Curved Surfaces: One-Step Immobilization of Human Erythrocyte Membranes on Silica Beads" , ChemPhysChem 4, 699 (2003).
[7] S. B. Rochal and V. L. Lorman, "Cytoskeleton Influence on Normal and Tangent Fluctuation Modes in the Red Blood Cells" , Phys. Rev. Lett. 96, 248102 (2006).
[8] L. C. L. Lin, N. Gov, and F. L. H. Brown, "Nonequilibrium membrane fluctuations driven by active proteins" , J. Chem. Phys. 124, 074903 (2006).
[9] R. D. Schaller, J. C. Johnson, and R. J. Saykally, "Time-Resolved Second Harmonic Generation Near-Field Scanning Optical Microscopy" , ChemPhysChem 4, 1243 (2003).
[10] J. P. Huang and K. W. Yu, J. "Nonlinear ac responses of electro-magnetorheological fluids" , Chem. Phys. 121, 7526 (2004).
[11] L. Gao, L. P. Gu, and Z. Y. Li, "Optical bistability and tristability in nonlinear metal/dielectric composite media of nonspherical particles" , Phys. Rev. E 68, 066601 (2003).
[12] T. B. Jones, Electromechanics of Particles (Cambridge University Press, New York, 1995).
[13] A. M. Woodward, A. Jones, and X. Zhang, "Rapid and non-invasive quantification of metabolic substrates in biological cell suspensions using non—linear dielectric spectroscopy with multivariate calibration and artificial neural networks. Principles and applications " , Bioelectrochem. Bioenerg. 40, 99 (1996).
[14] N. Gov, "Membrane Undulations Driven by Force Fluctuations of Active Proteins" , Phys. Rev. Lett. 93, 268104 (2004).
[15] R. D. Astumian, "Adiabatic Pumping Mechanism for Ion Motive ATPases" , Phys. Rev. Lett. 91, 118102 (2003).
[16] D. Nawarathna, J. H. Miller, Jr., J. R. Claycomb, G. Cardenas, and D. Warmflash, "Harmonic Response of Cellular Membrane Pumps to Low Frequency Electric Fields" , Phys. Rev. Lett. 95, 158103 (2005).
[17] T. Y. Tsong and R. D. Astumian, "863 -Absorption and conversion of electric field energy by membrane bound atpases" , Bioelectrochem. Bioenerg. 15, 457 (1986).
[18] R. D. Astumian and I. Derenyi, "Fluctuation driven transport and models of molecular motors and pumps" , Eur. Biophys. J. 27, 474 (1998).
[19] B. Chanda, O. K. Asamoah, R. Blunck, B. Roux, and F. Bezanilla, "Gating charge displacement in voltage-gated ion channels involves limited transmembrane movement" , Nature (London) 436, 852 (2005).
[20] C. L. Davey, H. M. Davey, and D. B. Kell, "On the dielectric properties of cell suspensions at high volume fractions" , Bioelectrochem. Bioenerg. 28, 319 (1992).
[21] A. M. Woodward and D. B. Kell, "On the nonlinear dielectric properties of biological systems Saccharomyces cerevisiae" , Bioelectrochem. Bioenerg. 24, 83 (1990).
[22] Garnett, J. C.M. Colors in metal glasses and in metal films [J]. Philos. Trans. R. Soc. London Ser. A., 1904, 203 (2): 385-420.
[23] Garnett, J. C. M. Absorptance to Selected Averaging Procedures and Solar Irradiance Distributions [J]. Trans. R. Soc. London Ser. A, 1906, 205 (5) : 237-284.
[24] Bruggeman, D. A. G. Berechnung verschiedener physikalischer Konstanten von heterogenen Substanzen [J]. Ann. Phys. (Leipzig), 1935, 24 (8) :636-679.
[25] L. D. Landau, E. M. Lifshitz, and L. P. Pitaevskii, Electrodynamics of Continuous Media, 2nd ed. (Pergamon, New York, 1984), Chap. II.
[26] J. Gimsa and D. Wachner, " A polarization model overcoming the geometric restrictions of the Laplace solution for spheroidal cells: obtaining new equations for field-induced forces and transmembrane potential" , Biophys. J. 77, 1316 (1999).
[27] M. Pavlin and D. Miklavcic, "Effective conductivity of a suspension of permeabilized cells: a theoretical analysis" , Biophys. J. 85, 719 (2003).
[28] J. P. Huang and K. W. Yu, "Enhanced nonlinear optical responses of materials: Composite effects" , Phys. Rep. 431, 87 (2006).
[29] D. A. G. Bruggeman, Ann. Phys. 「Leipzig(?) 24, 636 (1935).
[30] D. StroudandP. M. Hui, "Nonlinear susceptibilities of granular matter" , Phys. Rev. B 37, 8719 (1988).
[31] M. C. Fishman, "Genomics—zebrafish—the canonical vertebrate" , Science 294(5545), 1290-1291 (2001).
[32] G. Lomberk, "Animal models" , Pancreatol. 6(5), 427-428 (2006).
[33] J. R. Hove, R. W. Koster, A. S. Forouhar, G. Acevedo-Bolton, S. E. Fraser, and M. Gharib, "Intracardiac fluid forces are an essential epigenetic factor for embryonic cardiogenesis" , Nature (London) 421(6919), 172-178 (2003).
[34] A. S. Forouhar and M. Liebling, "The embryonic vertebrate heart tube is a dynamic suction pump" , Science 312(5774), 751-753 (2006).
[35] DJ; Jones, IL; Ellinor, PT, et al., "In vivo recording of adult zebrafish electrocardiogram and assessment of drug-induced QT prolongation Milan" AMERICAN JOURNAL OF PHYSIOLOGY-HEART AND CIRCULATORY PHYSIOLOGY. 291(1), H269-H273 (2006).
[36] Burns, CG; Milan, DJ; Grande, EJ, et al. "High-throughput assay for small molecules that modulate zebrafish embryonic heart rate" , NATURE CHEMICAL BIOLOGY 1(5) 263-264 (2005).
[37] W. R. Barrionuevo and W. W. Burggren, "02 consumption and heart rate in developing zebrafish (Danio rerio): influence of temperature and ambient O2" , Am. J. Physiol. Regulatory Integrative Comp. Physiol. 276, 505-513(1999).
[38] E: Jacob, M. Drexel, T. Schwerte, and B. Pelster, "Influence of hypoxia and of hypoxemia on the development of cardiac activity in zebrafish larvae" , Am. J. Physiol. Regulatory Integrative Comp. Physiol. 283, 911-917 (2002).
[39] C. G. Burns, D. J. Milan, E. J. Grande, W. Rottbauer, C. A. MacRae, and M. C. Fishman, "High-throughput assay for small molecules that modulate zebrafish embryonic heart rate" , Nat. Chem. Biol. 1(5), 263-264 (2005).
[40] M. Westerfield, The Zebrafish Book, Chap. 2, Eugene Press, Eugene, OR (1995).
[41] V. Viasnoff, F. Lequeux, and D. J. Pine, "Multispeckle diffusingwave spectroscopy: a tool to study slow relaxation and timedependent dynamics" , Rev. Sci. Instrum. 73(6), 2336-2344 (2002).
[42] A. V. Hallare, M. Schirling, T. Luckenbach, H. R. Kohler, and R. Triebskorn, "Combined effects of temperature and cadmium on developmental parameters and biomarker responses in zebrafish (Danio rerio) embryos", J. Therm. Biol. 30(1), 7-17 (2005). [43] Muller, M. ; Squier, J. A. ; Wilson, K. R. ; Brakenhoff, G. J. J. Microsc. 1998,191,266-74.
[44]Squier,J.A.;Muller,M.Appl.Opt.1999,38,5789-94.
[45]Richard D.Schaller,Justin C.Johnson,and Richard J.Saykally,"Nonlinear Chemical Imaging Microscopy;Near-Field Third Harmonic Generation Imaging of Human Red Blood Cells",Analytical Chemistry,72(21),5361-5364(2000).
[46]TSANG,T.Y.F,"OPTICAL 3RD-HARMONIC GENERATION AT INTERFACES",PHYSICAL REVIEW A,52(5) 4116-4125(1995).
[47]Barad,Y.;Eisenberg,H.;Horowitz,M.;Silberberg,Y.Appl.Phys.Lett.1997,70,922-4.
[48]Squier,J.A.;Muller,M.;Brakenhoff,G.J.;Wilson,K.R.Opt.Express 1998,3,315-24.
[49]Yelin,D.;Silberberg,Y.Opt.Express 1999,5,169-75.
[50]Yelin,D.;Silberberg,Y.;Barad,Y.;Patel,J.S.Appl.Phys.Lett:1999,74,3107-9.
© 2004-2018 中国地质图书馆版权所有 京ICP备05064691号 京公网安备11010802017129号 地址：北京市海淀区学院路29号 邮编：100083 电话：办公室：(+86 10)66554848；文献借阅、咨询服务、科技查新：66554700 |