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Issue 12, 2010
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Label-free molecular imaging of immunological synapses between dendritic and T cells by Raman micro-spectroscopy

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Abstract

Confocal Raman micro-spectroscopy (CRMS) was used to measure spectral images of immunological synapse formation between dendritic and T cells without using molecular labels or other invasive procedures. The purpose-built inverted CRMS instrument integrated an environmental enclosure and a near-infrared laser to allow measurements on live cells maintained under physiological conditions. The integration of the wide-field fluorescence also enabled viability assays and direct comparison between Raman spectral images and gold-standard immuno-fluorescence images for specific molecules. Raman spectral images of nucleus and proteins were built by fuzzy c-mean clustering method. The Raman images were found to be in good correspondence with the immuno-fluorescence images of DNA and actin. These results indicate that actin is a main contributor to the Raman spectrum of the cytoplasm of dendritic and T cells. While for control cells the Raman spectral images of proteins indicated a more homogeneous distribution of proteins in the cytoplasm of dendritic cells, they indicated a higher accumulation of proteins at the immunological synapses when dendritic cells were pre-treated with laminin. These conclusions were also supported by confocal immuno-fluorescence imaging after cell fixation and labelling. This study demonstrates the potential of CRMS for label-free non-invasive imaging of junctions between live cells. Therefore, this technique may become a useful tool for studying cellular processes in live cells and where non-invasive molecular specific imaging is desirable, such as cellcell interactions.

Graphical abstract: Label-free molecular imaging of immunological synapses between dendritic and T cells by Raman micro-spectroscopy

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Publication details

The article was received on 14 Jul 2010, accepted on 05 Oct 2010 and first published on 18 Oct 2010


Article type: Paper
DOI: 10.1039/C0AN00508H
Citation: Analyst, 2010,135, 3205-3212
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    Label-free molecular imaging of immunological synapses between dendritic and T cells by Raman micro-spectroscopy

    A. B. Zoladek, R. K. Johal, S. Garcia-Nieto, F. Pascut, K. M. Shakesheff, A. M. Ghaemmaghami and I. Notingher, Analyst, 2010, 135, 3205
    DOI: 10.1039/C0AN00508H

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