Combination of patch clamp and Raman spectroscopy for single-cell analysis.

In this contribution we present the combination of patch clamp with Raman spectroscopy for a label-free quantitative detection of intracellular components. Patch clamp is used to gain controlled access to the cytosol and internalize water-soluble compounds into the cell. The presence and concentration of these substances inside the living mammalian cell are probed by means of Raman spectroscopy in a label-free manner. A proof of principle was given using the carotinoid crocin as a sample compound that does not show specific interaction with the cell. When the intracellular crocin concentration as determined from the Raman spectra was monitored, the kinetics of internalization/diffusion into the cell could be characterized by a single-exponential function. Furthermore, the technique was successfully applied to observe differences in the internalization of free and protein-bound heme into the living cell. Although the peptide-capped microperoxidase MP-11 did not show specific interactions, free heme accumulated in the cell by binding to cellular components.