Surface-Enhanced Raman Scattering Dye-Labeled Au Nanoparticles for Triplexed Detection of Leukemia and Lymphoma Cells and SERS Flow Cytometry

Abstract

The labeling of cell surface receptors by fluorescent markers is an established method for the identification of cell phenotype in both research and clinical settings. Fluorescence dye labeling has inherent constraints, most notably the upper limit of labels per cell that may be probed using a single excitation source, in addition to a physical limit to the number of broad emission spectra that can be distinctly collected within the visible wavelength region. SERS labeling has the potential to mitigate these shortfalls. Herein, antibody-targeted, PEG-coated surface-enhanced Raman scattering (SERS) Au nanoparticles are used simultaneously to label three cell surface markers of interest on malignant B cells from the LY10 lymphoma cell line. The SERS probes were characterized by multiple methods to confirm their monodispersity and functionalization with both PEG and monoclonal antibodies. The specificity of the particles' cell labeling was demonstrated on both primary chronic lymphocytic leukemia and LY10 cells using SERS from cell suspensions and confocal Raman mapping, respectively. Fluorescence flow cytometry was employed to confirm the binding of SERS probes to LY10 over large cell populations, and the particles' SERS was collected directly from labeled cells using a commercial flow cytometer. To the best of our knowledge, this is the first demonstration of SERS flow cytometry from cells tagged with targeted SERS probes.

SERS Nanoparticles in Medicine: From Label-Free Detection to Spectroscopic Tagging

“…332 Later, MacLaughlin et al reported the use of SERS flow cytometry in identifying leukemia and lymphoma cells. 333 Here, 60 nm gold core SERS tags were conjugated to antibodies and used to detect three B cell surface markers (CD45, CD19, and CD20). Cell labeling was performed on two cell lines derived from patient samples, chronic lymphocytotic leukemia and LY10 cells.…”

Inorganic nanoparticles for biomedicine: where materials scientists meet medical research

“…3). Whilst only one cell type was detected in this study, three separate surface biomarkers could be simultaneously distinguished using Raman tags covalently bound to Au NPs [30]. An additional novelty in this study was the use of flow cytometry to differentiate between the different biomarkers.…”

“…SERS spectra of the three tags are shown in the corresponding spectra to the right. Flow cytometry can also be used to detect Raman tags (bottom graph), in this case malachite green (MGblack line), as shown by the shift in SERS intensity compared to Raman negative control (gray area)[30]. Copyright American Chemical Society, 2013.…”

Dielectrophoresis for Biomedical Sciences Applications: A Review

“…During the disease process, there are certain complex changes that occur in cells ranging from the molecular integrity to cell morphology [5]. There are various schemes used to detect diseases, including the use of dye marker labelling, enzyme-base labelling assays and nucleic acid-based assays [6,7,8,9,10]. All of these methods form the basis for nearly all of the pathology laboratory tests in hospitals.…”