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We also report on the time-dependent changes of cell nuclei as well as membrane receptor proteins during apoptosis analyzed by statistical multivariate methods.
An in-situ nondestructive characterization of the complicated chemical processes (like e.g. Here, we present the results of simultaneous and three-dimensional imaging of double organelles (nucleus and membrane) in single He La cells by means of either labelled or label-free surface-enhanced Raman spectroscopy (SERS).The CV-coated, CVa-coated, and MBA-coated Au NPs solutions displayed intense and uniform Raman response as shown in panels B to D of Fig. This is an important property of the Raman probes for cellular imaging application.It should be noted that the relatively low SERS intensity of label-free Au NPs revealed that NLS peptide and PAH ligands do not cause a significant Raman response during the detection of the intrinsic SERS signals from the cell components (Figure S2, black line).We show that representative and distinctly different intrinsic Raman signals of biomolecules from both membrane and nuclei can be selectively enhanced within single cells through membrane and nuclear-targeting label-free SERS probes, respectively.The Raman signals of two kinds of membrane targeting SERS probes and one nuclear-targeting SERS probe can be obtained from the same cell in three dimensions.After PAHylation of the dye-coated Au NPs, the Au NPs were subsequently modified with luteinizing hormone-releasing hormone (LHRH) and folate (FA) for membrane-targeting.
and NLS for nucleus-targeting via amide covalent linkages (Fig. In order to obtain the Raman spectra of biomolecules from the cell membrane and the nucleus without notable influence arising from the Raman reporters, the label-free targeting SERS Au NPs were designed as shown in Fig. The successful conjugation to PAH, NLS or FA, LHRH ligands was confirmed through an approx.Recently, nanoparticles (NPs) used as substrate for surface-enhanced Raman scattering (SERS) have attracted considerable attention as an emerging class of biolabels in cellular imaging, resulting in a high photostability of the SERS probes.Thirdly, an optimal contrast can be achieved by using red to near-infrared (NIR) excitation to minimize the disturbing autofluorescence of cells and tissues, making SERS an important tool for noninvasive imaging in living subjects.However, up to date, the goal using this technique to simultaneously and stereoscopically describe several subcellular organelles and biomolecules in single cells, and more importantly, to directly obtain the intrinsic chemical information of certain subcellular organelles has not been achieved to a satisfying extent.Here, we report the development of a new SERS strategy for triplex three-dimensional SERS imaging of a live cell, which allows for simultaneous SERS imaging of nucleus and membrane with high spatial resolution by means of confocal Raman microscopy.Furthermore, the two types of SERS probes (both labelled and label-free) will be used for the detection of apoptotic cells on single-cell level.