Highly multiparameter measurements are playing increasingly important roles in understanding complex cell systems. For fluorescence measurements using image and flow cytometry, the number of different probes that can be measured simultaneously is limited by the spectral width of the fluorophore emission spectra. Raman scattering spectra have much narrower spectral features, presenting the possibility of measuring many more probes within a given region of spectral space. To realize this potential, we have developed a Raman Flow Cytometer capable of measuring Raman scattering from single particles at high speeds. Nanoparticles exhibiting surface-enhanced Raman scattering (SERS) can serve as labels for antibodies or other binding probes, and offer the potential for higher levels of multiparameter and multiplexed measurements of cells and other particles than are possible with fluorescence alone.

Selected References:

A flow cytometer for the measurement of Raman spectra.
Watson, D.A., D.F. Gaskill, L.O. Brown, M.A. Naivar, S.W. Graves, S.K. Doorn, and J.P. Nolan (2008).
Cytometry 73A:119-128.

High throughput single nanoparticle spectroscopy.
David S Sebba, Dakota A Watson and John P Nolan (2009).
ACS Nano 2009; 3:1477-1484.

Spectral measurements of large particles by flow cytometry.
Watson, D.A., D.F. Gaskill, L.O. Brown, S.K. Doorn, J.P. Nolan (2009).
Cytometry 75:460-4.

Surface Enhanced Raman Scattering (SERS) Cytometry.
Nolan, J.P., DS Sebba (2011).
Methods in Cell Biology 102: 515-532.

Surface Enhanced Raman Scattering (SERS) Image Cytometry for High Content Screening.
Liu, E., J.P. Nolan (2014).
Advanced Flourescence Microscopy Techniques (PM Conn, ed), Elsevier.

Optimization of SERS tag intensity, binding footprint, and emittance.
Nolan, J.P., E. Duggan, D Condello (2014).
Journal of Bioconjugate Chemistry 25: 1233–1242.

Circulating tumor cell identification by functionalized silver-gold nanorods with super-enhanced SERS and photothermal resonances.
Nima Z.A., M. Mahmood, Y. Xu, T. Mustafa, F. Watanabe, D.A. Nedoskin, M.A. Juratli, T. Fahmi, E.I. Galanzha, J.P Nolan, A. Basnakian, V.P. Zharov, A.S. Biris (2014).
Scientific Reports 4:4752.