An evaluation of the application of the aperture infrared SNOM technique to biomedical imaging

Asingle human oesophageal adenocarcinoma cell (OE33) has been imaged using aperture infrared scanning near-field optical microscopy (IR-SNOM) in transmission and reflection and also by Fourier-transform infrared (FTIR) microspectroscopy in transmission only. This work presents the first images obtained in both transmission and reflection of the same specimen using the aperture IR-SNOM technique. The results have been used to compare the two SNOM modes and also the two techniques, which have complementary capabilities. The SNOM technique necessitates a very stable source and a careful choice of wavelengths, since it is too slow to yield images at the thousands of wavelengths obtained with FTIR. However the SNOM technique is not diffraction limited and with careful fabrication of tips can yield images with high spatial resolution. There is no significant correlation between the SNOM images obtained in transmission and reflection and the correlations between images obtained at different wavelengths vary with the different imaging modes. These results are attributed to the strong dependence of the evanescent wave on both the wavelength and the distance between the tip and the source of the signal within the sample. While both transmission and reflection SNOM images show some correlation with topography this is not a dominant effect. These results indicate that with suitable calibration a combination of reflection and transmission aperture IR-SNOM measurements has the potential to reveal information on the depth distribution of the chemical structure of a specimen.