Identification of site-specific isotopic labels by vibrational spectroscopy in the electron microscope

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Science  01 Feb 2019:
Vol. 363, Issue 6426, pp. 525-528
DOI: 10.1126/science.aav5845

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Mapping isotopically labeled alanine

Electron microscopy of organic materials must avoid the destructive effects of electron beam impact. One approach is to measure vibrational spectra with electron energy-loss spectroscopy in a mode where the electron beam grazes the sample and couples to it through evanescent modes. Hachtel et al. used such methods to probe carbon-12– and carbon-13–labeled alanine crystals, which exhibited an isotopic shift in the asymmetric carbon-oxygen stretching mode. They used this property to map the distribution of labeled clusters of alanine on length scales of tens of nanometers.

Science, this issue p. 525


The identification of isotopic labels by conventional macroscopic techniques lacks spatial resolution and requires relatively large quantities of material for measurements. We recorded the vibrational spectra of an α amino acid, l-alanine, with damage-free “aloof” electron energy-loss spectroscopy in a scanning transmission electron microscope to directly resolve carbon-site–specific isotopic labels in real space with nanoscale spatial resolution. An isotopic red shift of 4.8 ± 0.4 milli–electron volts in C–O asymmetric stretching modes was observed for 13C-labeled l-alanine at the carboxylate carbon site, which was confirmed by macroscopic infrared spectroscopy and theoretical calculations. The accurate measurement of this shift opens the door to nondestructive, site-specific, spatially resolved identification of isotopically labeled molecules with the electron microscope.

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