WST-8 Assay for Living Cell Viability Detection by Surface-enhanced Raman Spectroscopy†

doi:10.7503/cjcu20150282

Abstract

Abstract:

Surface-enhanced Raman scattering(SERS) technique was used to develop and optimize a novel and quantitative 2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium monosodium salt(WST-8) assay for living cell viability. The influences of pH and dosage of reductant nicoti-namide adenine dinucleotide(NADH) on WST-8 formazan signal were examined. SERS signals of formazan with high resolution and great diversity between WST-8 and formazan were observed in neutral conditions. Meanwhile, the signals of formazan(560, 1351, 1598, 1616 cm^-1) could be found clearly when the concentration of NADH was four times that of WST-8. Compared with the traditional method, this assay appears higher sensitivity and selectivity for WST-8 formazan. Moreover, the detection limit of formazan is as low as 10 μmol/L.

Key words: 2-(2-Methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2, 4-disulfophenyl)-2H-tetrazolium monosodium salt(WST-8), Surface-enhanced Raman scattering(SERS), Cell viability detection

CLC Number:

O657.37

TrendMD:

YANG Jin, LIU Zhuo, WANG Xiaohui, SUI Huimin, MAO Zhu, ZHAO Bing. WST-8 Assay for Living Cell Viability Detection by Surface-enhanced Raman Spectroscopy^†[J]. Chem. J. Chinese Universities, 2015, 36(10): 1900.

Figures/Tables 8

Fig.1 TEM image of silver collid

Fig.2 UV-Vis absorption spectra(A) and SERS spectra(B, λex=633 nm) of 1-Methoxy PMS(a), NADH(b), WST-8 formazan(c) and WST-8(d) solutions

Fig.3 UV-Vis spectra of the mixture of WST-8, 1-Methoxy PMS and NADH at different time(A) and the corresponding I460 nm-t curve(B)

Fig.4 UV-Vis absorption spectra(A) and SERS spectra(B, λex=633 nm) of WST-8 at different pH values

Fig.5 UV-Vis absorption spectra(A) and SERS spectra(B, λex=633 nm) of WST-8 formazan at different pH valuesa. WST-8 formazan+NaOH(pH=13.12); b. WST-8 formazan+HCl(pH=0.57); c. WST-8 formazan(pH=7.35).

Fig.6 UV-Vis absorption spectra(A) and SERS spectra(B, λex=633 nm) of WST-8 solution with different concentrations of NADHc(NADH)/c(WST-8): (A) a—e. 0, 1∶2, 1∶1, 3∶2, 2∶1; (B) a—e. 0, 1∶1, 2∶1, 3∶1, 4∶1.

Fig.7 UV-Vis spectra(A) and SERS spectra(B, λex=633 nm) of WST-8 stock solution at different concentrations(A) c(WST-8)/(mmol·L-1): a. 0.0125; b. 0.025; c. 0.05; d. 0.075; e. 0.10.(B) c(WST-8)/(μmol·L-1): a—j. 1.00; 0.50; 0.33; 0.25; 0.20; 0.17; 0.14; 0.13; 0.11; 0.10.

Fig.8 UV-Vis spectra(A) and SERS spectra(B, λex=633 nm) of WST-8 formazan stock solution at different concentrationsc(WST-8 formazan)/(μmol·L-1): a—k. 0, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100.

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