Immunoassay of Rat Brain Tissue by Surface-enhanced Raman Spectroscopy†

doi:10.7503/cjcu20150635

Abstract

Abstract:

Surface-enhanced Raman scattering(SERS) was used to develop and optimize a novel, quick and simple immunohistochemical ABC(avidin-biotin-HRP complex) assay for detecting specific skeleton protein neurofilament-200(NF-200) in the frozen sections of rat brain tissue. In this study, anti mouse NF-200 labeled colloid gold was used as immunoprobe, which was added into the frozen sections of rat brain tissue containing NF-200 protein. Intense SERS signals of p-mercaptobenzoic acid(MBA) at 1076 and 1589 cm^-1 were observed clearly after the specific binding of antigen and antibody. Meanwhile, the SERS spectra and the immunohistochemical staining effect of antigen and antibody complex with different concentrations of anti NF-200 were examined. Compared with the traditional method, this assay can simplify the procedure, moreover, it showed higher sensitivity and selectivity.

Key words: Neurofilament-200(NF-200), Surface-enhanced Raman scattering(SERS), Immunohisto-chemistry

CLC Number:

O657.37

TrendMD:

YANG Jin, LIU Zhuo, SU Hongyang, PENG Ru, SONG Wei, ZHAO Bing. Immunoassay of Rat Brain Tissue by Surface-enhanced Raman Spectroscopy^†[J]. Chem. J. Chinese Universities, 2015, 36(11): 2317.

Figures/Tables 6

Fig.1 TEM image of AuNPs

Fig.2 UV-Vis absorption spectra of AuNPs@Ab-MBA@BSA(a), AuNPs@MBA(b), AuNPs@Ab-MBA(c) and AuNPs(d)

Fig.3 SERS spectra(λex=633 nm)of AuNPs@MBA(a), AuNPs@Ab-MBA(b), AuNPs@Ab-MBA@BSA(c), AuNPs@Ab-MBA@BSA-tissue(d), AuNPs-tissue(e), AuNPs@Ab(f)

Fig.4 Optical images of NF-200 immunoreactivity in mice brain tissue slice(A) and liver tissue slice(B) The nerve cell is labeled in square frame.

Fig.5 SERS spectra(λex=633 nm)of AuNPs@Ab-MBA@BSA-Tissue with different concentrations of anti NF-200 Concentration of anti NF-200/(μg·mL-1): a. 1.000; b. 0.500; c. 0.250; d. 0.125; e. 0.0625.

Fig.6 Optical images of NF-200 immunoreactivity in mice brain tissue slice with different concentrations of anti NF-200

References 27

[1]	Gu L. J., Jian Z. H., Stary C., Xiong X. X., Neurochem. Res., 2015, 40(9), 1786—1791
[2]	Zhao M. L., Zhang L. Z., Wang Z. Y., Wang X. L., Wang Y. Y., Wei H. Y., Li R. H., Du Y. F., Int. Health, 2015, 7(4), 293—297
[3]	Friess S. H., Lapidus J. B., Brody D. L., J. Neurotrauma, 2015, 32(11), 791—800
[4]	Hwang I. K., Koh U. S., Lee J. C., Yoo K. Y., Song J. H., Jung J. Y., Nam Y. S., Lee I. S., Kang T. C., Won M. H., J. Neurol. Sci., 2005, 239(1), 59—66
[5]	Liang W.H., Zhang W.W., Zhao S.F., Li Q. N., Liang H., Ceng R. C., Neurol. Sci.,36(5), 707—712
[6]	Jia Y. J., Wu D., Zhang R. P.,Shuang W. B., Sun J. P., Hao H. H., An Q. J., Liu Q., Neurosci. Lett., 2014, 573, 46—51
[7]	Chen M. H., Liu C., Cai J.Y.,Song X. J., Li X. W., Sun J., Acta Medica Mediterr., 2014, 30(1), 95—99
[8]	Hsu S. M., Raine L., Fanger H., J. Histochem. Cytochem., 1981, 29(4), 577—580
[9]	Zlatian O. M., Comanescu M. V., Rosu A. F., Rosu L., Cruce M., Gaman A. E., Calina C. D., Sfredel V., Rom. J. Morphol. Embryo, 2015, 56(1), 175—181
[10]	Goodpaster T., Randolph H.J., J. Histochem. Cytochem., 2014, 62(3), 197—204
[11]	Garamvoelgyi R., Repa I., Lorincz B., Jakab C., Balka G., Lelovics Z., Magy. Allatorvosok, 2013, 135(9), 535—542
[12]	Endo M., Molohashi A., Akahira J.I., Akagami Y., Minamiya Y., Virchows Arch., 2013, 463(2), 325—335
[13]	Anderson J. J., Fordham S., Overman L., Dignum H., Wood K., Proctor S. J., Crosier S., Angus B., Culpin R. E., Mainou F. T., Int. J. Oncol., 2009, 35(5), 961—971
[14]	Lunedo S. N., Thomaz S. V., Castro E. A., Telles J. E. Q., Acta Histochem., 2012, 114(3), 252—258
[15]	Rivera P. M., Osuna A. K. P., Dominguez A. S., Garza R. G., Int. J. Morphol., 2011, 29(2), 575—580
[16]	Stevenson R., Stokes R. J., MacMillan D., Armstrong D., Faulds K., Wadsworth R., Kunuthur S., Suckling C. J., Graham D., Analyst, 2009, 134(8), 1561—1564
[17]	Zhang B. H., Wang H. S., Lu L. H., Ai K. L., Zhang G., Cheng X. L., Adv. Funct. Mater., 2008, 18(16), 2348—2355
[18]	Chen L., Xue X. X., Cui Y. C., Han X. X., Xu W. Q., Zhao B., Chem. J. Chinese Universities, 2015, 36(8), 1505—1510
	(陈雷, 薛向欣, 崔运成, 韩晓霞, 徐蔚青, 赵冰. 高等学校化学学报, 2015, 36(8), 1505—1510)
[19]	Hering K., Cialla D., Ackermann K., Dörfer T., Möller R., Schneidewind H., Fritzsche W., Popp J., Anal. Bioanal. Chem., 2008, 390, 113—124
[20]	Shanmukh S., Jones L., Zhao Y. P., Driskell J., Tripp R., Dluhy R., Anal. Bioanal. Chem., 2008, 390, 1551—1555
[21]	Yang W., Gooding J. J., Hibbert D. B., J. Electroanal. Chem., 2001, 516, 10—16
[22]	Xu S. P., Ji W. Q., Xu W. Q., Li X. L., Wang L. Y., Bai Y. B., Zhao B., Ozaki Y., Analyst, 2004, 129(1), 63—68
[23]	Lin C. C., Yang Y. M., Chen Y. F., Yang T. S., Chang H. C., Biosens. Bioelectron., 2008, 24(2), 178—183
[24]	Chon H., Lee S., Son S. W., Oh C. H., Choo J. B., Anal. Chem., 2009, 81(8), 3029—3034
[25]	Sun L., Yu C. X., Irudayaraj J., Anal. Chem., 2007, 79(11), 3981—3988
[26]	Dey P. , Olds W., Blakey I., Thurecht K. J., Izake E. L., Fredericks P. M., J. Raman Spectrosc., 2013, 44(12), 1659—1665
[27]	Lee P. C., Meisel D., J. Phys. Chem., 1982, 86, 3391—3395