Noninvasive imaging of protein metabolic labeling in single human cells using stable isotopes and Raman microscopy

H.J. van Manen, Aufrid T.M. Lenferink, Cornelis Otto

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84 Citations (Scopus)

Abstract

We have combined nonresonant Raman microspectroscopy and spectral imaging with stable isotope labeling by amino acids in cell culture (SILAC) to selectively detect the incorporation of deuterium-labeled phenylalanine, tyrosine, and methionine into proteins in intact, single HeLa cells. The C−D stretching vibrational bands in these amino acids are observed in the 2100−2300 cm−1 spectral region that is devoid of vibrational contributions from other, nondeuterated intracellular constituents. We found that incubation with deuterated amino acids for 8 h in cell culture already led to clearly detectable isotope-related signals in Raman spectra of HeLa cells. As expected, the level of isotope incorporation into proteins increased with incubation time, reaching 55% for deuterated phenylalanine after 28 h. Raman spectral imaging of HeLa cells incubated with deuterium-labeled amino acids showed similar spatial distributions for both isotope-labeled and unlabeled proteins, as evidenced by Raman ratio imaging. The SILAC−Raman methodology presented here combines the strengths of stable isotopic labeling of cells with the nondestructive and quantitative nature of Raman chemical imaging and is likely to become a powerful tool in both cell biology applications and research on tissues or whole organisms
Original languageUndefined
Pages (from-to)9576-9582
Number of pages7
JournalAnalytical chemistry
Volume80
Issue number24
DOIs
Publication statusPublished - 2008

Keywords

  • METIS-254098
  • IR-75806

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