Combining Immunolabeling and Surface-Enhanced Raman Spectroscopy on Cell Membranes

被引:57
|
作者
Hodges, Matthew D. [1 ]
Kelly, Jemma G. [2 ]
Bentley, Adam J. [1 ]
Fogarty, Simon [1 ]
Patel, Imran I. [2 ]
Martin, Francis L. [2 ]
Fullwood, Nigel J. [1 ]
机构
[1] Univ Lancaster, Sch Hlth & Med, Div Biomed & Life Sci, Lancaster LA1 4YQ, England
[2] Univ Lancaster, Lancaster Environm Ctr, Ctr Biophoton, Lancaster LA1 4YQ, England
基金
英国生物技术与生命科学研究理事会;
关键词
cell membrane; surfaced-enhanced Raman; silver nanoparticles; immunolabeling; keratan sulfate; lipids; proteins; IN-VIVO; IMMUNO-PCR; SCATTERING; NANOPARTICLES; SPECTRA; SERS; MANIPULATION; TAGS;
D O I
10.1021/nn202652h
中图分类号
O6 [化学];
学科分类号
0703 ;
摘要
We applied surface-enhanced Raman spectroscopy (SERS) to immunolabeled endothelial cells to derive enhanced spectra of the biomolecular makeup of the cellular surface. A two-step immunolabeling protocol with gold-conjugated antibodies coupled with silver enhancement to attach silver nanoparticles to the cell surface was employed. This approach generated similar to 50-fold SERS enhancement of spectral signals. The SERS spectra exhibited several SERS-enhanced peaks associated with cell membrane components. The SERS detection of silver nanoparticles proved more far more sensitive than conventional light microscopy techniques. The SERS enhancement allowed us to carry out spectral mapping using wavenumbers associated with membrane components that correlated directly with the distribution of silver nanoparticles. SERS has the potential to detect immunolabeling at lower levels than Is possible using conventional immunolabeling methods while simultaneously providing unique, spatially defined, biochemical information.
引用
收藏
页码:9535 / 9541
页数:7
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