Functional imaging of a single cell: far-field infrared super-resolution microscopy using autofluorescence detection

Tsutomu Ohmori; Keiichi Inoue; Makoto Sakai; Masaaki Fujii; Miya Ishihara M.D.; Makoto Kikuchi M.D.

doi:10.1117/12.808903

23 February 2009 Functional imaging of a single cell: far-field infrared super-resolution microscopy using autofluorescence detection

Tsutomu Ohmori, Keiichi Inoue, Makoto Sakai, Masaaki Fujii, Miya Ishihara M.D., Makoto Kikuchi M.D.

Author Affiliations +

Proceedings Volume 7182, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues VII; 71820G (2009) https://doi.org/10.1117/12.808903
Event: SPIE BiOS, 2009, San Jose, California, United States

Abstract

We demonstrated cell imaging without any stain by far-field 2-color infrared (IR) super-resolution microscopy, combining laser fluorescence microscope and picosecond transient fluorescence detected IR (TFD-IR) spectroscopy. TFD-IR spectroscopy detects IR absorption by monitoring fluorescence due to an electronic transition from a vibrational excited level by an additional visible light. By using the IR microscopy based on TFD-IR spectroscopy, the spatial resolution of the image can be increased to the visible diffraction limit of sub-μm, i.e., the IR is super-resolved. Cell auto-fluorescence due to flavin molecules was monitored for label-free detection of the cellular components. The fluorescence image of an A549 cell was obtained by introducing both an IR light at 3300 nm and a visible light at 560 nm. The spatial resolution of the image was estimated to be 1.6 μm. This is about 2.5-times higher resolution than the diffraction limit of IR light. The fluorescence intensity of the images at 3448 nm was smaller than that at 3300 nm, corresponding to the smaller IR absorption. Therefore, IR spectral imaging of a single cell was achieved with superresolution.

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Tsutomu Ohmori, Keiichi Inoue, Makoto Sakai, Masaaki Fujii, Miya Ishihara M.D., and Makoto Kikuchi M.D. "Functional imaging of a single cell: far-field infrared super-resolution microscopy using autofluorescence detection", Proc. SPIE 7182, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues VII, 71820G (23 February 2009); https://doi.org/10.1117/12.808903

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